CN109554485A - It is a kind of for Non-invasive detection fetal chromosomal to be measured whether be aneuploid kit and its application specific probe group - Google Patents

Abstract

The invention discloses it is a kind of for Non-invasive detection fetal chromosomal to be measured whether be aneuploid kit and its application specific probe group.Probe groups include 500 specific probes.Each specific probe includes DNA fragmentation 2.The nucleotide sequence of the DNA fragmentation 2 of 500 specific probes successively such as the sequence of sequence table 1 from 5 ' ends 16-93 to shown in sequence 500 16-93 from 5 ' ends of sequence table.It is demonstrated experimentally that kit provided by the invention can detecte whether No. 13 chromosomes of fetus, No. 18 chromosomes, No. 21 chromosomes and sex chromosome to be measured are aneuploid, testing result and amniotic fluid gold mark testing result are completely the same.The present invention has important application value.

Description

It is a kind of for Non-invasive detection fetal chromosomal to be measured whether be aneuploid kit And its application specific probe group

Technical field

The invention belongs to medical domains, and in particular to whether one kind is non-multiple for Non-invasive detection fetal chromosomal to be measured The kit and its application specific probe group of body, more particularly to it is a kind of for detecting No. 13 chromosomes of fetus to be measured, No. 18 chromosomes, 21 Number chromosome and sex chromosome whether be aneuploid kit and its application specific probe group.

Background technique

Birth defect is to endanger one of the three big diseases of mankind's disease health.According to the latest data of the World Health Organization, the whole world Prevalence rate of birth defect be about 3%-6%, just have the newborn of nearly 8,000,000 birth defect every year.Chromosome abnormality is to make The main reason at a large amount of birth defects.The most common chromosome disease normally behaves as chromosomal DNA and repeats and lack, And chromosomal aneuploidy, the latter refer to anomaly existing or lacking for complete chromosome.When organism having less than or More than normal diploid chromosome quantitative when, a large amount of off-notes will be generated and cause a variety of syndromes.Down syndrome, Also known as 21- patau syndrome is the most common example of chromosomal aneuploidy, including a No. 21 additional chromosomes.It is other Common chromosomal aneuploidy has 13- patau syndrome, E trisomy, Turner syndrome and Klinefelter comprehensive Sign.

Pre-natal diagnosis chromosome aneuploid generally uses the screening of serology biochemistry and villus acquisition, amniocentesis, bleeding of the umbilicus The invasive diagnostic methods such as puncture.Amniocentesis is the goldstandard of diagnosis, but traumatic strong, and with risk of miscarriage.Serology Screening is despite damage-free type, but recall rate and false positive rate are lower.

1997, Lu Yu penetrating judgment, which is beheaded, time had found fetus dissociative DNA (cfDNA), but technology at that time is difficult logarithm Huge cfDNA is measured effectively to be detected.Until 2007, second generation sequencing technologies (NGS) occurred, and researcher just has found The research tool of one " claiming hand ".Meanwhile scientist readily recognizes that, the high pass flow characteristic of second generation sequencing technologies is also very suitable Run the noninvasive prenatal gene detection of exhibition jointly.Directly to genome sequencing, there are at high cost, data analysis complexity, period length etc. to ask Topic.Target sequence capture sequencing reduces sequencing cost, improves sequencing depth, can more precisely find region sequence undetermined Arrange the information changed.

A kind of novel fetal chromosomal disease detection skill that NIPT (noninvasive Prenatal Screening) occurs as antenatal detection field Art passes through high pass for relatively traditional chorionic villi sampling or amniocentesis etc. have the antenatal detection method of " invasive " It measures sequencing technologies and " Noninvasive " antenatal detection is carried out to the fetus dissociative DNA in mother's peripheral blood, more safely, quickly, quasi- Really, existing Prenatal Screening, pre-natal diagnosis system are effectively supplemented.

Summary of the invention

It whether is aneuploid the purpose of the present invention is detecting fetal chromosomal to be measured, so that whether prenatal diagnosis of fetal suffers from There is corresponding prenatal diagnosis.

The present invention protect first it is a kind of for detect No. 13 chromosomes of fetus to be measured, No. 18 chromosomes, No. 21 chromosomes and Sex chromosome whether be aneuploid kit, it may include probe groups；

The probe groups may include 500 specific probes；Each specific probe may include DNA fragmentation 2；The DNA fragmentation 2 It can be the nucleotide sequence of No. 13 chromosome, the nucleotide sequence of No. 18 chromosome, the nucleotide sequence of No. 21 chromosomes, X dye A part in the nucleotide sequence of colour solid or the nucleotide sequence of Y chromosome.

In mentioned reagent box, the DNA fragmentation 2 can be 60-140 (such as 60-78,78-100,100-140,60 A, 78,100 or 140) nucleotide composition single strand dna.

In mentioned reagent box, the nucleotide sequence of the DNA fragmentation 2 of 500 specific probes successively can be such as sequence table Sequence 1 from 5 ' ends the 16-93 sequences 500 to sequence table from 5 ' ends shown in 16-93.

In mentioned reagent box, each specific probe may also include DNA fragmentation 1 and DNA fragmentation 3；The DNA fragmentation 1 is located at 5 ' ends of the DNA fragmentation 2；The DNA fragmentation 3 is located at 3 ' ends of the DNA fragmentation 2.

The DNA fragmentation 1 and the DNA fragmentation 3 can for 10-20 (such as 10-15,15-20,10,15 or 20) nucleotide composition single strand dna.

Each specific probe from 5 ' ends to 3 ' ends successively may include the DNA fragmentation 1, the DNA fragmentation 2 and described DNA fragmentation 3.

Any of the above-described probe groups can be specifically made of 500 specific probes.Each specific probe is from 5 ' ends to 3 ' End can be specifically made of the DNA fragmentation 1, the DNA fragmentation 2 and the DNA fragmentation 3.

Any of the above-described kit may also include primer pair first；The primer pair first can be made of primer 1 and primer 2； The primer 1 may include the DNA fragmentation 1；The primer 2 may include the DNA fragmentation 3.

The primer 1 concretely DNA fragmentation 1.

The primer 2 concretely DNA fragmentation 3.

The primer 1 and/or the primer 2 can have biotin labeling.

Any of the above-described DNA fragmentation 1 and/or any of the above-described DNA fragmentation 3 have biotin labeling (i.e. in reagent Without containing under conditions of primer pair first in box, biotin mark directly can be carried out to the DNA fragmentation 1 and/or the DNA fragmentation 3 Note).

The nucleotide sequence of any of the above-described DNA fragmentation 1 can be as shown in the sequence 501 of sequence table.

The nucleotide sequence of any of the above-described DNA fragmentation 3 can be as shown in the sequence 502 of sequence table.

The nucleotide sequence of any of the above-described 500 specific probes successively can be such as sequence 1 to the sequence table of sequence table Shown in sequence 500.

Any of the above-described kit can be specifically made of the probe groups.

Any of the above-described kit can be specifically made of the probe groups and the primer pair first.

Any of the above-described kit may also include connector 1 (being used to prepare pregnant woman blood plasma dissociative DNA to be measured library), connector 2 (being used to prepare pregnant woman blood plasma dissociative DNA to be measured library), primer 3 (for closing the connector 1), primer 4 are (for closing State connector 2), primer 5 (for PCR amplification) and primer 6 (being used for PCR amplification)；

The connector 1 from 5 ' ends to 3 ' ends successively may include DNA fragmentation a (i.e. sequence measuring joints sequence 1), DNA fragmentation b With DNA fragmentation c；3 ' the ends of the DNA fragmentation c can be T；

The connector 2 from 5 ' ends to 3 ' ends successively may include DNA fragmentation d and DNA fragmentation e；

The DNA fragmentation b and DNA fragmentation d can be made of at random A, T, C and G；

3 ' end reverse complementals of the 5 ' ends of the DNA fragmentation e and the DNA fragmentation a；

The primer 5 may include the DNA fragmentation A；

The primer 3 from 5 ' ends to 3 ' ends successively may include the DNA fragmentation A, DNA fragmentation B and DNA fragmentation C；Institute 5 ' the ends for stating DNA fragmentation C and the DNA fragmentation a are completely the same；The DNA fragmentation B can be made of at random A, T, C and G；

The primer 6 may include the DNA fragmentation D；

The primer 4 from 5 ' ends to 3 ' ends successively may include the DNA fragmentation D, DNA fragmentation E and DNA fragmentation F；Institute 3 ' the ends for stating DNA fragmentation F and the DNA fragmentation e are completely the same；The DNA fragmentation E can be made of at random A, T, C and G.

The DNA fragmentation b and the DNA fragmentation d may be the same or different.

The length of the DNA fragmentation c concretely 1bp.

The length of the DNA fragmentation b and the DNA fragmentation d be 6-14bp (such as 6-8bp, 8-14bp, 6bp, 8bp or 14bp)。

The length of the DNA fragmentation A is 17-25bp (such as 17-21bp, 21-25bp, 17bp, 21bp or 25bp).

The length of the DNA fragmentation B is 1-8bp (such as 1-4bp, 4-8bp, 1bp, 4bp or 8bp).

The length of the DNA fragmentation D is 17-25bp (such as 17-21bp, 21-25bp, 17bp, 21bp or 25bp).

The length of the DNA fragmentation E is 10-25bp (such as 10-16bp, 16-25bp, 10bp, 16bp or 25bp).

The primer 4 may also include Barcode flag sequence；The Barcode flag sequence is located at the DNA fragmentation E (in one embodiment of the invention, Barcode flag sequence is located in the DNA fragmentation E upstream, downstream or centre Between).The length of the Barcode flag sequence is 6-14bp (such as 6-8bp, 8-14bp, 6bp, 8bp or 14bp).It is described Barcode flag sequence can be made of at random A, T, C and G, for distinguishing different samples to be tested when sequencing data processing.

3 ' ends of the connector 1, the primer 5 and the primer 6 can carry out thio-modification.

5 ' ends of the connector 2 can carry out phosphorylation modification.

The connector 1 from 5 ' ends to 3 ' ends specifically can by DNA fragmentation a (sequence measuring joints sequence 1), DNA fragmentation b and DNA fragmentation c composition.In an embodiment of the present invention, it is sequenced using illumina platform.Correspondingly, DNA fragmentation a (sequencing Joint sequence 1) nucleotide sequence (from 5 ' to 3 ') can be ACACTCTTTCCCTACACGACGCTCTTCCGATCT.According to survey The difference of sequence platform can also be substituted for the sequence measuring joints sequence being made of other nucleotide sequences.The nucleotide of the connector 1 Sequence (from 5 ' to 3 ') specifically can be as shown in sequence 507 in sequence table.

The connector 2 can be specifically made of from 5 ' ends to 3 ' ends DNA fragmentation d and DNA fragmentation e.In reality of the invention It applies in example, is sequenced using illumina platform.Correspondingly, the nucleotides sequence of the DNA fragmentation e (sequence measuring joints sequence 2) Arranging (from 5 ' to 3 ') can be AGATCGGAAGAGCACACGTCT.According to the difference of microarray dataset, can also be substituted for by other cores The sequence measuring joints sequence of nucleotide sequence composition.The nucleotide sequence (from 5 ' to 3 ') of the connector 2 specifically can be such as sequence in sequence table Shown in column 508.

The primer 3 specifically can be by the DNA fragmentation A, the DNA fragmentation B and the DNA piece from 5 ' ends to 3 ' ends Section C composition.The nucleotide sequence (from 5 ' to 3 ') of the primer 3 specifically can be as shown in sequence 503 in sequence table.

The primer 4 specifically can be by the DNA fragmentation D, the DNA fragmentation E (containing described from 5 ' ends to 3 ' ends Barcode flag sequence) and DNA fragmentation F composition.The nucleotide sequence (from 5 ' to 3 ') of the primer 4 specifically can be such as sequence In list shown in sequence 504.

The primer 5 can be specifically made of the DNA fragmentation A.The nucleotide sequence (from 5 ' to 3 ') of the primer 5 is specific It can be as shown in sequence 505 in sequence table.

The primer 6 can be specifically made of the DNA fragmentation D.The nucleotide sequence (from 5 ' to 3 ') of the primer 5 is specific It can be as shown in sequence 506 in sequence table.

Any of the above-described kit specifically can be by the probe groups, the connector 1, the connector 2, the primer 3, institute Primer 4, the primer 5 and the primer 6 is stated to form.

Any of the above-described kit specifically can be by the probe groups, the primer pair first, the connector 1, the connector 2, the primer 3, the primer 4, the primer 5 and the primer 6 form.

Any of the above-described kit may also include the carrier for recording judgment criteria first and judgment criteria second.

The judgment criteria first be judge pregnant woman fetus to be measured target autosome number be increase, reduce or just Often；The target autosome can be No. 13 chromosomes, No. 18 chromosomes or No. 21 chromosomes.

The judgment criteria first can be as follows: first calculating pregnant woman's target autosome Z-score to be measured according to formula (1)；

ChrN_Z-score: pregnant woman's target autosome z-score to be measured；

%ChrN_sample: the reads ratio of pregnant woman's target autosome capture region to be measured；

Mean ChrN reference: target autosome capture region reads ratio is averaged in reference database Value；

S.D.%ChrN reference: the standard of target autosome capture region reads ratio in reference database Variance；

Then it makes the following judgment: if target autosome Z-score >=3 of pregnant woman to be measured, pregnant woman fetus to be measured Target autosome number increase；If the target autosome Z-score < 3 of -3 < pregnant woman to be measured, pregnant woman's tire to be measured The target autosome number of youngster is normal；If the target chromosome Z-score of pregnant woman to be measured≤- 3, pregnant woman fetus to be measured Target autosome number is reduced；If target autosome number increases or target autosome number is reduced, fetus mesh Mark autosome is aneuploid (for example, No. 13 chromosome number increase will cause 13- patau syndrome, No. 18 dyeing Body number increase will cause E trisomy, and No. 21 chromosome number increase will cause 21- patau syndrome (i.e. Tang Shi Syndrome))；If target autosome number is normal, fetus target autosome is euploid.

Fetus target autosome is aneuploid, that is, suffers from target autosome aneuploid disease.

The judgment criteria second is to judge that the sex chromosome number of pregnant woman fetus to be measured is to increase, reduce or normal.

The judgment criteria second can be as follows:

(1) sex of foetus is first judged according to the reads ratio of pregnant woman's Y chromosome capture region to be measured: if pregnant woman Y to be measured contaminates Colour solid capture region reads ratio > 0.001, then sex of foetus is male；If pregnant woman's Y chromosome capture region reads to be measured Ratio≤0.001, then sex of foetus is women.

(2) if fetus is women, the X chromosome Z-score of pregnant woman to be measured is first calculated according to formula (2)；

ChrX_Z-score: pregnant woman's X chromosome z-score to be measured；

%ChrN_sample: the reads ratio of pregnant woman's X chromosome capture region to be measured；

Mean ChrN reference: the average value of X chromosome capture region reads ratio in female's fetal data library；

S.D.%ChrN reference: in female's fetal data library the standard side of X chromosome capture region reads ratio Difference；

Then make the following judgment: if X chromosome Z-score >=3 of pregnant woman to be measured, the X of pregnant woman fetus to be measured contaminates Colour solid number increases；If the X chromosome Z-score < 3 of -3 < pregnant woman to be measured, the X chromosome number of pregnant woman fetus to be measured Normally；If the X chromosome Z-score of pregnant woman to be measured≤- 3, the X chromosome number of pregnant woman fetus to be measured is reduced；It is fetal Not Wei women when, if X chromosome number increase or X chromosome number reduce, fetus X chromosome be aneuploid (for example, X chromosome number increase will cause poly-x female syndrome；The reduction of X chromosome number will cause Turner syndrome)；If X chromosome Number is normal, then fetus X chromosome is euploid.

When sex of foetus is women, fetus X chromosome is aneuploid, that is, suffers from X chromosome aneuploid disease.

(3) if fetus is male, the X chromosome Z- of pregnant woman to be measured is obtained first, in accordance with following steps (a)-(c) Score:

(a) normal male database X chromosome and Y chromosome ratio mean value are calculated according to formula (3):

C: normal male database X chromosome and Y chromosome ratio mean value are (in one embodiment of the invention, through counting It calculates, 2.817) normal male database X chromosome and Y chromosome ratio mean value are；

%ChrX_MDB: the average value of X chromosome capture region reads ratio in normal male database；

%ChrY_MDB: the average value of Y chromosome capture region reads ratio in normal male database.

(b) the reads ratio of converted rear pregnant woman's X chromosome capture region to be measured is calculated according to formula (4):

%ChrX is the reads ratio of converted rear pregnant woman's X chromosome capture region to be measured；

%ChrX_sampleFor the reads ratio of pregnant woman's X chromosome capture region to be measured；

C is normal male database X chromosome and Y chromosome ratio mean value；

%ChrY_sampleFor the reads ratio of pregnant woman's Y chromosome capture region to be measured；

(c) pregnant woman's X chromosome Z-score to be measured is calculated according to formula (5):

ChrX_Z-sco_re: pregnant woman's X chromosome z-score to be measured；

%ChrX: the reads ratio of pregnant woman's X chromosome capture region to be measured after converted；

Mean ChrXreference: the average value of X chromosome capture region reads ratio in female's fetal data library；

S.D.%ChrX reference: in female's fetal data library the standard side of X chromosome capture region reads ratio Difference；

Then it makes the following judgment: if X chromosome Z-score >=3 of pregnant woman to be measured and pregnant woman's Y chromosome to be measured capture Give a birth out in the reads ratio < normal pregnancies of region male fetus Y chromosome capture region reads ratio average value, then to The increase of X chromosome number, the Y chromosome number for surveying pregnant woman fetus are normal；If X chromosome Z-score >=3 of pregnant woman to be measured and It gives a birth out in pregnant woman's Y chromosome capture region reads ratio >=normal pregnancies to be measured the Y chromosome capture region of male fetus The average value of reads ratio, then the Y chromosome number increase of pregnant woman fetus to be measured, X chromosome number are normal；If -3 < are waited for The X chromosome Z-score < 3 of pregnant woman is surveyed, then the X chromosome of pregnant woman fetus to be measured and Y chromosome number are normal；If to be measured The X chromosome Z-score of pregnant woman≤- 3, then the X chromosome number of pregnant woman fetus to be measured is reduced, the normal (feelings of Y chromosome number Condition is generally not in, because male does not have X chromosome that can not then survive)；When sex of foetus is male, if X chromosome number (the reduced number of situation of X chromosome is generally not in, because male does not have X chromosome for mesh increase or the reduction of X chromosome number Can not then survive), Y chromosome number it is normal, then fetus X chromosome be aneuploid (such as: the increase of X chromosome number can make At Klinefelter syndrome)；If Y chromosome number increases, X chromosome number is normal, fetus Y chromosome is non-multiple Body (such as: Y chromosome number increase will cause XYY syndrome)；If X chromosome and Y chromosome number are normal, tire Youngster's X chromosome and Y chromosome are euploid；

The reference database can be by the data of the healthy pregnant women of M healthy fetus of giving a birth and the data of N healthy males It collectively constitutes；M and N is 50 or more natural number；

The normal male database can be made of the data of H healthy males；H can be 30 or more natural number；

Can be given birth to by the R status data of healthy pregnant women of female fetus of female's fetal data library form；R can be 30 or more Natural number；

Person under test can be healthy pregnant women, healthy male or the healthy pregnant women for female fetus of giving a birth out of childbirth healthy fetus；It obtains The step of obtaining the data of person under test can be as follows:

(K1) person under test's plasma DNA library is prepared；It can be described for preparing the connector in person under test's plasma DNA library Connector 1 and the connector 2；

(K2) after completing step (K1), person under test's plasma DNA library is hybridized with the probe groups, is obtained To hybrid product；

(K3) after completing step (K2), target sequence is collected from the hybrid product, is obtained target sequence and is captured library；

(K4) the target sequence capture library obtained step (K3) carries out high-flux sequence, obtains sequencing initial data；

(K5) the sequencing initial data for obtaining step (K4) cuts off joint sequence, low quality base, and filtering is (such as filtering length Degree is less than 40bp sequence), it then compares to the corresponding position with reference to genome hg19；

(K6) after completing step (K5), the repetitive sequence due to caused by PCR amplification preference is removed, the number of the person under test is obtained According to.

When sex of foetus is male, fetus X chromosome is aneuploid, Y chromosome is euploid, that is, suffers from X chromosome Aneuploid disease；Fetus Y chromosome is aneuploid, X chromosome is euploid, that is, suffers from Y chromosome aneuploid disease.

In one embodiment of the invention, any of the above-described reference database specifically can be by the healthy tire of 400 childbirths The data of the healthy pregnant women of youngster and the data of 50 healthy males collectively constitute.Any of the above-described normal male database can be by The data composition of 50 healthy males.It any of the above-described female's fetal data library can be pregnant by the health of 400 healthy fetus of giving a birth Give a birth out in woman female fetus pregnant woman data composition.

In one embodiment of the invention, in 400 normal pregnancies (healthy pregnant women of i.e. 400 childbirth healthy fetus) The average value of Y chromosome capture region reads ratio of male fetus of giving a birth out is 0.00345.

Any of the above-described kit specifically can be by the probe groups, the connector 1, the connector 2, the primer 3, institute State primer 4, the primer 5, the primer 6 and the carrier composition for recording judgment criteria first and judgment criteria second.

Any of the above-described kit specifically can be by the probe groups, the primer pair first, the connector 1, the connector 2, the primer 3, the primer 4, the primer 5, the primer 6 and judgment criteria first and the judgment criteria second of recording Carrier composition.

Any of the above-described kit may also include the conventional reagent for carrying out PCR amplification.

Any of the above-described kit may also include the conventional reagent that DNA library is hybridized with probe.

The present invention also protect it is a kind of detect fetal chromosomal to be measured whether be aneuploid method, successively may include as follows Step:

(1) pregnant woman blood plasma dissociative DNA to be measured library is prepared；

(2) after completing step (1), the pregnant woman blood plasma dissociative DNA to be measured library and the progress of any of the above-described probe groups is miscellaneous It hands over, obtains hybrid product；

(3) after completing step (2), target sequence is collected from the hybrid product, is obtained target sequence and is captured library；

(4) after completing step (3), target sequence capture library is sequenced, tire to be measured is judged according to sequencing data Whether No. 13 chromosomes, No. 18 chromosomes, No. 21 chromosomes and the sex chromosome of youngster is aneuploid.

In the step (1), the connector for preparing pregnant woman blood plasma dissociative DNA to be measured library can be any of the above-described connector 1 With the connector 2.

Described the step of preparing pregnant woman blood plasma dissociative DNA library to be measured is concretely: extracting pregnant woman blood plasma trip to be measured first From DNA, then preparing pregnant woman blood plasma dissociative DNA to be measured library using KAPA Hyper library construction Kit, (connector can be upper State any connector 1 and the connector 2).The KAPA Hyper library construction Kit concretely KAPA The product of Biosystems company, article No. KK8504.

In the step (2), " pregnant woman blood plasma dissociative DNA to be measured library is hybridized with the probe groups, is hybridized The step of product ", can be as follows:

(2-1) prepares enrichment system；The enrichment system includes the pregnant woman blood plasma dissociative DNA to be measured library, the spy Needle group, the primer 3 and the primer 4；

(2-2) takes the enrichment system, and 60-70 DEG C of (such as 60-65 DEG C, 65-70 DEG C, 60 DEG C, 65 DEG C or 70 DEG C) hybridization is rich Collect 16h or more (such as 16h), obtains hybrid product.

In the step (2-1), the enrichment system can be by pregnant woman blood plasma dissociative DNA to be measured library, people cot-1DNA, salmon Milt DNA, the primer 3, the primer 4, probe groups and hybridization buffer (65 DEG C of preheatings) composition.

In the step (2-1), enrichment system specifically can be by 48 μ L pregnant woman blood plasma dissociative DNA to be measured library described in 100 μ L (the pregnant woman blood plasma dissociative DNA to be measured containing 1 μ g), 12 μ L harvesting buffers, 5 μ L probe groups and 35 μ L hybridization buffers (65 DEG C of preheatings) Composition.The harvesting buffer can be by 3.5 parts by volume people cot-1DNA aqueous solutions, 3.5 parts by volume salmon sperm dna aqueous solutions and 3 Parts by volume primer mixed liquor (being made of primer 3, primer 4 and water) forms.In harvesting buffer, the concentration of people cot-1DNA can be 30%-45% (such as 30%-35%, 35%-45%, 30%, 35%, 40% or 45%) (v/v), the concentration of salmon sperm dna can For 5%-25% (such as 5%-15%, 15%-25%, 5%, 10%, 15%, 20% or 25%), the concentration of primer 3 can be 0.2- 1nmol/ μ L (such as 0.2-0.5nmol/ μ L, 0.5-1nmol/ μ L, 0.2nmol/ μ L, 0.3nmol/ μ L, 0.5nmol/ μ L, 0.6nmol/ μ L or 1nmol/ μ L), the concentration of primer 4 can be 0.2-1nmol/ μ L (such as 0.2-0.5nmol/ μ L, 0.5-1nmol/ μ L, 0.2nmol/ μ L, 0.3nmol/ μ L, 0.5nmol/ μ L, 0.6nmol/ μ L or 1nmol/ μ L).

The hybridization buffer can be for containing 1.0-1.5M (such as 1.0-1.25M, 1.25-1.5M, 1M, 1.25M or 1.5M) NaCl, 0.1-0.3M (such as 0.1-0.15M, 0.15-0.3M, 0.1M, 0.125M, 0.15M, 0.2M or 0.3M) sodium citrate, 0.08-0.12g/100mL (such as 0.08-0.10g/100mL, 0.10-0.12g/100mL, 0.08g/100mL, 0.10g/100mL Or 0.12g/100mL) BSA and 5-9% (such as 5-7%, 7-9%, 5%, 6%, 7%, 8% or 9%) (v/v) Tween20 water Solution.

In the step (2-2), the hybridization can be carried out in PCR instrument.Response procedures can are as follows: 95 DEG C of 7min, 65 DEG C of 16h with On.

In the step (3), the step of " collecting target sequence from hybrid product, obtain target sequence and capture library ", can be such as Under:

(3-a) collects the hybrid product with magnetic bead；

The product that (3-b) is collected using step (3-a) is template, using the primer pair of the primer 5 and the composition of the primer 6 PCR amplification is carried out, pcr amplification product is target sequence capture library.

The step (3-a) specifically may include following step (3-1), step (3-2), step (3-3) and step (3-4).

The step (3-b) specifically may include following step (3-5), step (3-6) and step (3-7).

In the step (3), the step of " collecting target sequence from hybrid product, obtain target sequence and capture library ", specifically may be used It is as follows:

(3-1) prepares the solution of Streptavidin coating magnetic bead；

(3-2) takes centrifuge tube, and the solution and the hybrid product of the Streptavidin coating magnetic bead is added, is incubated for；So It is placed on magnetic frame or centrifugation, abandons supernatant；

After (3-3) completes step (3-2), the centrifuge tube is taken, wash buffer, washing is added；

After (3-4) completes step (3-3), the centrifuge tube is taken, magnetic bead is resuspended；It is subsequently placed in magnetic frame or centrifugation, is collected Supernatant；

(3-5) prepares PCR amplification system；PCR amplification system includes supernatant, the claim 7 to 10 that step (3-4) is collected Described in primer 5 and the primer 6；

(3-6) takes the PCR amplification system, carries out PCR amplification, obtains pcr amplification product；

(3-7) takes the pcr amplification product, purifying, obtains target sequence, i.e. target sequence captures library.

In the step (3-2), the incubation can for incubation at room temperature 0.5-2h (such as 0.5-1h, 1-2h, 0.5h, 1h or 2h).It is placed in magnetic frame and is concretely placed in magnetic frame 1min.

In the step (3-3), " wash buffer, washing is added " can be divided into the progress of two steps: drift is added in the first step Wash buffer 1 washs；Second step is added wash buffer 2 and washs.Purpose is by the spy not in conjunction with Streptavidin MagneSphere Different probe is washed off.

The wash buffer 1 can be the sodium citrate buffer solution containing 0.05-0.2% (m/v) SDS (as contained 0.05- The sodium citrate buffer solution of 0.1% (m/v) SDS, contains 0.05% (m/ at the sodium citrate buffer solution containing 0.1-0.2% (m/v) SDS V) sodium citrate buffer solution of SDS, the sodium citrate buffer solution containing 0.1% (m/v) SDS or the lemon for containing 0.2% (m/v) SDS Sour sodium buffer).Sodium citrate buffer solution can be the aqueous solution of NaCl containing 175g/L and 88g/L trisodium citrate；PH value is 7.4。

The wash buffer 2 concretely contains the sodium citrate buffer solution dilution of 0.05-0.2% (m/v) SDS (such as Sodium citrate buffer solution dilution containing 0.05-0.1% (m/v) SDS, the buffered sodium citrate containing 0.1-0.2% (m/v) SDS Liquid dilution, the sodium citrate buffer solution dilution containing 0.05% (m/v) SDS, the buffered sodium citrate for containing 0.1% (m/v) SDS Liquid dilution or the sodium citrate buffer solution dilution for containing 0.2% (m/v) SDS).Sodium citrate buffer solution dilution can be 1 body Product part sodium citrate buffer solution and 9 parts by volume water mix.

In the step (3-4), " magnetic bead is resuspended " is enriched with eluent resuspension by the way that library is added.The library enrichment elution Liquid can be the NaOH aqueous solution that concentration is 0.1M.

In the step (3-5), the PCR amplification system can by step (3-4) collect supernatant, neutralization buffer and PCR reaction solution composition.

The Tris-HCl buffer of the neutralization buffer concretely pH7.5,1M.

PCR reaction solution by Phusion Hot Start II DNA Polymerase, dNTP, DMSO, primer 5, primer 6, MgCl₂, KCl, Nonidet P40 and water composition.

3mL PCR reaction solution can be specifically 0.05U/ μ L Phusion Hot Start II DNA by 100 μ L concentration Polymerase (product of thermo company, article No. F549S) and 2900 μ L PCR mixbuffer composition.PCR Mixbuffer is dATP containing 0.2mM, 0.2mM dTTP, 0.2mM dCTP, 0.2mM dGTP, 5% (v/v) DMSO, 2.5pmol Primer 5,2.5pmol primer 6,4mMMgCl₂, 500mM KCl and 0.8% (v/v) Nonidet P40 aqueous solution.

In the step (3-6), the program of described " carrying out PCR amplification " can are as follows: 98 DEG C of 30sec；98 DEG C of 30sec, 65 DEG C 30sec, 72 DEG C of 30sec, 15 circulations；72 DEG C of 5min, 4 DEG C of ∞.

In the step (3-7), the purifying can be to utilize Agencourt AMPure XP Nucleic acid purification kits (product of Beckman Coulter company, article No. A63881) is purified.

In the step (4), " target sequence capture library is sequenced, tire to be measured is judged according to sequencing data fruit Whether No. 13 chromosomes, No. 18 chromosomes, No. 21 chromosomes and the sex chromosome of youngster is aneuploid " it can be by the target sequence Capture library is sequenced, and sequencing initial data is obtained；Sequencing initial data is handled, sequencing data is obtained；According to survey Ordinal number is it is judged that whether No. 13 chromosomes, No. 18 chromosomes, No. 21 chromosomes and the sex chromosome of fetus to be measured are aneuploid.

It is described will sequencing initial data carry out processing include the following steps:

(4-1) will be sequenced initial data and cut off joint sequence, low quality base, and (such as Filter length is less than 40bp sequence for filtering Column), it then compares to the corresponding position with reference to genome hg19；

After (4-2) completes step (4-1), the repetitive sequence due to caused by PCR amplification preference is removed, sequencing data is obtained.

In the step (4), the judgement mark of " judging whether fetal chromosomal to be measured is aneuploid according to sequencing data " It will definitely be any of the above-described judgment criteria first and/or any of the above-described judgment criteria second.

The present invention also protects target sequence to capture library, is made of the target sequence that any of the above-described the method obtains.

The present invention also protects any of the above-described probe groups.

The present invention also protects the application of any of the above-described probe groups, can be W1) or W2):

W1) detect whether No. 13 chromosomes of fetus, No. 18 chromosomes, No. 21 chromosomes and sex chromosome to be measured are non-multiple Body；

W2 No. 13 chromosomes of fetus, No. 18 chromosomes, No. 21 chromosomes and sex chromosome to be measured) are detected.

The platform of any of the above-described sequencing can be Nextseq500, X Ten or NovaSeq.

It is demonstrated experimentally that kit provided by the invention can detecte No. 13 chromosomes of fetus to be measured, No. 18 chromosomes, No. 21 Whether chromosome and sex chromosome are aneuploid, testing result and amniotic fluid gold mark testing result (belonging to invasive pre-natal diagnosis) It is completely the same.The present invention has important application value.

Specific embodiment

Embodiment below facilitates a better understanding of the present invention, but does not limit the present invention.

Experimental method in following embodiments is unless otherwise specified conventional method.

Test material as used in the following examples is unless otherwise specified to buy from routine biochemistry reagent shop It arrives.

Pregnant woman in following embodiments is to the equal informed consent of experiment content.

The nucleotide sequence of primer involved in following embodiments is shown in Table 1.

Table 1

Note:^*G indicates that G carries out thio-modification,^*A indicates that A carries out thio-modification,^*T is that T carries out thio-modification, and P is phosphorylation Modification, N A, T, C or G.

Embodiment 1, for Non-invasive detection fetal chromosomal to be measured whether be aneuploid kit preparation

One, the preparation of probe groups

1, according to the specific regions of No. 13 chromosome of people, the specific regions of No. 18 chromosome of people, people No. 21 dyeing Specific regions, the specific regions of people's X chromosome and the specific regions of people's Y chromosome of body design and synthesize 500 probes. Each probe is made of 108 nucleotide, successively includes DNA fragmentation 1, DNA fragmentation 2 and DNA piece from 5 ' ends to 3 ' ends Section 3.The nucleotides sequence of DNA fragmentation 1 is classified as 5 '-GACTACATGGGACAT-3 '.The nucleotides sequence of DNA fragmentation 3 is classified as 5 '- GGAACCTACGACGTA-3'.DNA fragmentation 2 is made of 78 nucleotide.DNA fragmentation 2 is the special of No. 13 chromosome of people Region, the specific regions of No. 18 chromosome of people, the specific regions of No. 21 chromosome of people, people's X chromosome specific regions Or a part of the specific regions of people's Y chromosome.

2, after completing step 1, biotin labeling is carried out to probe.Specific steps conduct: 500 probes are uniformly mixed Total volume is the ddH of 1.2mL₂In O, take wherein 15 μ L using general PCR primer to (by primer 1:5 '-bio- GACTACATGGGACAT-3 ' (bio indicates that 5 ' ends have biotin labeling) and primer 2: 5 '-GGAACCTACGACGTA-3 ' Composition) PCR amplification (three Guan Jinhang can be divided) is carried out, obtain pcr amplification product.

3, pcr amplification product is taken, with MinElute PCR Purification Kit (product of Qiagen company, article No. 28006) to be purified, purified product is obtained；500ng purified product is taken later, with MyOne streptavidin magnetic bead (product of Invitrogen company, article No. 35602) is combined；NaOH solution processing is added later, purpose is will not have The complementary strand denaturation of biotin labeling elutes；Later by 100 DEG C of the formamide liquid scrubbing of entire magnetic bead, make probe It is separated from magnetic bead；Ethanol precipitation is finally used, the probe groups of biotin labeling are obtained.

Two, for Non-invasive detection fetal chromosomal to be measured whether be aneuploid kit preparation

It whether is the kit of aneuploid by harvesting buffer, Hybridization Buffer for Non-invasive detection fetal chromosomal to be measured Liquid, library combination buffer, wash buffer 1, wash buffer 2, library are enriched with eluent, neutralization buffer, PCR reaction solution With the probe groups composition of step 1 preparation.

Harvesting buffer is by 3.5 parts by volume people cot-1DNA aqueous solutions, 3.5 parts by volume salmon sperm dna aqueous solutions and 3 volumes Part primer mixed liquor (being made of primer 3, primer 4 and water) forms.In harvesting buffer, the concentration of people cot-1DNA is 35% (v/v), the concentration of salmon sperm dna is 15% (v/v), and the concentration of primer 3 is 0.5nmol/ μ L, and the concentration of primer 4 is 0.5nmol/μL。

Hybridization buffer: NaCl containing 1.25M, 0.125M sodium citrate, 0.1g/100mL BSA and 7% (v/v) The aqueous solution of Tween20.

Library combination buffer: the Tris-HCl buffer of pH7.5,10mM of NaCl containing 1M and 1mM EDTA.

Wash buffer 1: contain the sodium citrate buffer solution of 0.1% (m/v) SDS.Sodium citrate buffer solution is containing 175g/L The aqueous solution of NaCl and 88g/L trisodium citrate；PH value is 7.4.

Wash buffer 2: contain the sodium citrate buffer solution dilution of 0.1% (m/v) SDS.Sodium citrate buffer solution dilution Liquid is that 1 parts by volume sodium citrate buffer solution and 9 parts by volume water mix.

Library is enriched with eluent: concentration is the NaOH aqueous solution of 0.1M.

Neutralization buffer: the Tris-HCl buffer of pH7.5,1M.

3mL PCR reaction solution is 0.05U/ μ L Phusion Hot Start II DNA by 100 μ L concentration Polymerase (product of thermo company, article No. F549S) and 2900 μ L PCR mixbuffer composition.PCR Mixbuffer is dATP containing 0.2mM, 0.2mM dTTP, 0.2mM dCTP, 0.2mM dGTP, 5% (v/v) DMSO, 2.5pmol Primer 5,2.5pmol primer 6,4mMMgCl₂, 500mM KCl and 0.8% (v/v) Nonidet P40 aqueous solution.

Whether the noninvasive antenatal detection fetus to be measured of kit prepared by embodiment 2, embodiment 1 suffers from chromosome aneuploid The foundation of the method for disease

The present inventor passes through many experiments, establishes the noninvasive antenatal detection of kit prepared using embodiment 1 The method whether fetus to be measured suffers from prenatal diagnosis.Specific step is as follows:

One, pregnant woman blood plasma dissociative DNA to be measured extracts

1, maternal blood 10mL to be measured is acquired with the anticoagulant blood vessel of EDTA, 4 DEG C, 1600g centrifugation 15min collect supernatant 1 (being placed in low adsorption centrifuge tube).

2, after completing step 1, the supernatant 1 is taken, 12000rpm is centrifuged 5min, collects supernatant 2 and (is placed in low adsorption centrifugation Pipe).Supernatant 2 is blood plasma.

3, after completing step 2,1mL supernatant 2 is taken, with large volume free nucleic acid extracts kit (Tiangeng biochemical technology (north Capital) Co., Ltd product, article No. DP710) extract plasma DNA (cfDNA).The cfDNA is named as pregnant woman to be measured Plasma DNA.

Two, the preparation in pregnant woman blood plasma dissociative DNA to be measured library

The pregnant woman blood plasma dissociative DNA to be measured for taking 5ng step 1 to obtain, utilizes KAPA Hyper library construction Kit (product of KAPA Biosystems company, article No. KK8504) prepares pregnant woman blood plasma dissociative DNA to be measured library.It prepares to be measured The connector in pregnant woman blood plasma dissociative DNA library is single stranded DNA 1 and single stranded DNA 2.

Pregnant woman blood plasma dissociative DNA to be measured library is taken, Qubit is carried out and is quantitatively detected with 2% agarose gel electrophoresis.

Three, the building in target sequence capture library

1, enrichment system is prepared.Enrichment system is 100 μ L, (is waited for containing 1 μ g by 48 μ L pregnant woman blood plasma dissociative DNA to be measured library Survey pregnant woman blood plasma dissociative DNA), 12 μ L harvesting buffers, 5 μ L probe groups and 35 μ L hybridization buffers (65 DEG C of preheatings) composition.

2, after completing step 1, the enrichment system is taken, being placed in PCR instrument progress overnight hybridization enrichment, (hybridization time is at least 16h), enriched product is obtained.

Response procedures: 95 DEG C of 7min, 65 DEG C of 16h or more.

3, Streptavidin coating magnetic bead solution acquisition (Streptavidin be coated with magnetic bead save liquid in save, into The purpose of this step of row is that removal saves liquid)

(1) low adsorption centrifuge tube is taken, 50 μ L are addedM-280 Streptavidin MagneSphere, is subsequently placed in magnetic force Frame 1min.

(2) after completing step (1), supernatant is abandoned, 50 library μ L combination buffers are added, magnetic bead, then of short duration centrifugation is resuspended.

(3) after completing step (2), the low adsorption centrifuge tube is taken, is placed in magnetic frame 1min, supernatant is abandoned, 50 μ is then added Magnetic bead is resuspended in the library L combination buffer.

(4) after completing step (3), the low adsorption centrifuge tube is taken, is placed in magnetic frame 1min, supernatant is abandoned, 50 μ is then added Magnetic bead is resuspended in the library L combination buffer, obtains the solution of Streptavidin coating magnetic bead.

4, it is incubated for capture altogether

(1) after completing step 3, the low adsorption centrifuge tube is taken, whole enriched products that step 2 obtains are added, are vortexed 5sec is shaken, 1h on the rotation blending instrument of room temperature is subsequently placed in.

(2) after completing step (1), the low adsorption centrifuge tube is taken, is placed in magnetic frame 1min, abandons supernatant.

5, washing and PCR amplification

(1) after completing step 4, the low adsorption centrifuge tube is taken, 500 μ L wash buffers 1 are added, mixes, is placed in rotation Blending instrument rotates 15min.

(2) after completing step (1), the low adsorption centrifuge tube is taken, is placed on magnetic frame and stands 2min, abandon supernatant.

(3) after completing step (2), the low adsorption centrifuge tube is taken, 500 μ L wash buffers 2 (65 DEG C of preheatings) are added, 65 DEG C of incubation 10min.

(4) after completing step (3), the low adsorption centrifuge tube is taken, is placed on magnetic frame and stands 2min, abandon supernatant.

(5) step (4) are repeated twice.

Carry out above-mentioned steps purpose be will not withThe probe that M-280 Streptavidin MagneSphere combines is washed Fall.

(6) after completing step (5), the low adsorption centrifuge tube is taken, 30 libraries μ L are added and are enriched with eluent, magnetic bead is resuspended, It is placed in room temperature rotation blending instrument 10-20min.

(7) after completing step (6), the low adsorption centrifuge tube is taken, is placed on magnetic frame and stands 1min.

(8) after completing step (7), supernatant is transferred to centrifuge tube, 40 μ L neutralization buffers and 30 μ L PCR are then added Reaction solution, pipettor suction play mixing and are placed on PCR instrument progress PCR amplification, obtain pcr amplification product.

PCR program: 98 DEG C of 30sec；98 DEG C of 30sec, 65 DEG C of 30sec, 72 DEG C of 30sec, 15 circulations；72 DEG C of 5min, 4 DEG C ∞。

6, it purifies

After completing step 5, the pcr amplification product is taken, Agencourt AMPure XP Nucleic acid purification kits are utilized (product of Beckman Coulter company, article No. A63881) purifying, obtains amplified library purified product.

The library amplification purification product is target sequence capture library.

Four, high-flux sequence and data analysis

1, the target sequence capture library obtained step 3 is sequenced by two generations such as Nextseq500, X Ten, NovaSeq Platform carries out high-flux sequence, obtains sequencing initial data.

2, complete step 1 after, will sequencing initial data with cutadapt software (network address are as follows: https: // Cutadapt.readthedocs.io/en/stable/ joint sequence, low quality base (mass value is less than 10), filtering) are cut off Length is less than 40bp sequence, then by bwa software (network address are as follows: http://bio-bwa.sourceforge.net/) or Bowtie software (http://http://bowtie-bio.sourceforge.net/bowtie2/) is compared to reference to genome The corresponding position of hg19.

3, after completing step 2, with biobambam2 software (https: //software.broadinstitute.org/ Gatk/ the repetitive sequence due to caused by PCR amplification preference) is removed, the data of pregnant woman to be measured is obtained, further obtains pregnant woman to be measured The capture region reads ratio of target chromosome.

4, the foundation of reference database

(1) person under test's plasma DNA extracts

" pregnant woman to be measured " in step 1 is replaced with into certain person under test, other steps are constant, obtain person under test's blood plasma trip From DNA.

(2) preparation in person under test's plasma DNA library

The person under test's plasma DNA for taking 5ng step (1) to obtain respectively, utilizes KAPA Hyper library construction Kit Prepare person under test's plasma DNA library.The connector for preparing pregnant woman blood plasma dissociative DNA to be measured library is single stranded DNA 1 and single-stranded DNA2。

(3) building in target sequence capture library

" pregnant woman blood plasma dissociative DNA to be measured library " in step 3 is replaced with to person under test's blood plasma of step (2) preparation respectively Dissociative DNA library, other steps are constant, obtain target sequence and capture library.

(4) the target sequence capture library obtained step (3) is surveyed by two generations such as Nextseq500, X Ten, NovaSeq Sequence platform carries out high-flux sequence, obtains sequencing initial data.

(5) the sequencing initial data cutadapt software for obtaining step (4) cuts off joint sequence, low quality base (mass value is less than 10), Filter length are less than 40bp sequence, are then compared by bwa software or bowtie software to reference gene The corresponding position of group hg19.

(6) after completing step (5), the repetitive sequence due to caused by PCR amplification preference is removed with biobambam2 software, is obtained To the data of the person under test.

Person under test totally 450, respectively 400 normal pregnancies (healthy pregnant women for healthy fetus of giving a birth) and 50 male Property.

The data of 450 persons under test collectively constitute reference database.

The data of 50 male persons under test collectively constitute normal male database.

The data of person under test of female fetus of giving a birth out in 400 normal pregnancies collectively constitute female's fetal data library.

The data of person under test of male fetus of giving a birth out in 400 normal pregnancies collectively constitute male fetal data library.

5, target chromosome (No. 13 chromosome, No. 18 chromosome, No. 21 chromosome, X in reference database are calculated Chromosome or Y chromosome) capture region reads ratio average value (Mean) and standard variance (SD:Standrad Deviation)。

Target chromosome (No. 13 chromosome, No. 18 chromosome, No. 21 chromosome, X chromosome in reference database Or Y chromosome) capture region reads ratio average value (Mean) and standard variance (SD:Standrad Deviation) Value is shown in Table 2.

Table 2

	The average value of capture region reads ratio	The standard variance of capture region reads ratio
			No. 13 chromosome	0.070480234	0.003783135
No. 18 chromosome	0.056481519	0.004469963
			No. 21 chromosome	0.053474884	0.003796539
X chromosome	0.138318218944452(M)/0.140807034370486(F)	0.0115653390816059(M)/0.0119558674493583(F)
			Y chromosome	0.00345129816422603(M)/-	0.00128275369533018(M)/-

6, after completing step 5, pregnant woman's target autosome Z-score to be measured is first calculated according to the following equation；

ChrN_Z-sco_re: pregnant woman's target autosome z-score to be measured；

%ChrN_sample: the reads ratio of pregnant woman's target autosome capture region to be measured；

Mean ChrN reference: target autosome capture region reads ratio is averaged in reference database Value；

S.D.%ChrN reference: the standard of target autosome capture region reads ratio in reference database Variance.

Then it makes the following judgment: if target autosome Z-score >=3 of pregnant woman to be measured, pregnant woman fetus to be measured Target autosome number increase；If the target autosome Z-score < 3 of -3 < pregnant woman to be measured, pregnant woman's tire to be measured The target autosome number of youngster is normal；If the target chromosome Z-score of pregnant woman to be measured≤- 3, pregnant woman fetus to be measured Target autosome number is reduced.

Target autosome number increases or the reduction of target autosome number will cause prenatal diagnosis. For example, No. 13 chromosome number increase will cause 13- patau syndrome, No. 18 chromosome number increase will cause 18- tri- Body syndrome, No. 21 chromosome number increase will cause 21- patau syndrome (i.e. Down syndrome).

7, after completing step 5, judge that the sex chromosome number of pregnant woman fetus to be measured is to increase, reduce or normal.Specifically Steps are as follows:

(1) sex of foetus is first judged according to the reads ratio of pregnant woman's Y chromosome capture region to be measured: if pregnant woman Y to be measured contaminates Colour solid capture region reads ratio > 0.001, then sex of foetus is male；If pregnant woman's Y chromosome capture region reads to be measured Ratio≤0.001, then sex of foetus is women.

(2) if fetus is women, the X chromosome Z-score of pregnant woman to be measured is first calculated according to the following equation；

ChrX_Z-score: pregnant woman's X chromosome z-score to be measured；

%ChrN_sample: the reads ratio of pregnant woman's X chromosome capture region to be measured；

Mean ChrN reference: the average value of X chromosome capture region reads ratio in female's fetal data library；

S.D.%ChrN reference: in female's fetal data library the standard side of X chromosome capture region reads ratio Difference.

Then make the following judgment: if X chromosome Z-score >=3 of pregnant woman to be measured, the X of pregnant woman fetus to be measured contaminates Colour solid number increases；If the X chromosome Z-score < 3 of -3 < pregnant woman to be measured, the X chromosome number of pregnant woman fetus to be measured Normally；If the X chromosome Z-score of pregnant woman to be measured≤- 3, the X chromosome number of pregnant woman fetus to be measured is reduced.

When sex of foetus is women, X chromosome number increases or the reduction of X chromosome number will cause chromosome aneuploidy Body disease.X chromosome number increase will cause poly-x female syndrome.The reduction of X chromosome number will cause Turner syndrome.

(3) if fetus is male, the X chromosome Z- of pregnant woman to be measured is obtained first, in accordance with following steps (a)-(c) Score:

(a) normal male database X chromosome and Y chromosome ratio mean value are calculated according to the following equation:

C: normal male database X chromosome and Y chromosome ratio mean value；

%ChrX_MDB: the average value of X chromosome capture region reads ratio in normal male database；

%ChrY_MDB: the average value of Y chromosome capture region reads ratio in normal male database.

It is computed, normal male database X chromosome and Y chromosome ratio mean value are 2.817.

(b) the reads ratio of converted rear pregnant woman's X chromosome capture region to be measured is calculated according to the following equation:

%ChrX is the reads ratio of converted rear pregnant woman's X chromosome capture region to be measured；

%ChrX_sampleFor the reads ratio of pregnant woman's X chromosome capture region to be measured；

C is normal male database X chromosome and Y chromosome ratio mean value；

%ChrY_sampleFor the reads ratio of pregnant woman's Y chromosome capture region to be measured.

(c) pregnant woman's X chromosome Z-score to be measured is calculated according to the following equation:

ChrX_Z-sco_re: pregnant woman's X chromosome z-score to be measured；

%ChrX: the reads ratio of pregnant woman's X chromosome capture region to be measured after converted；

Mean ChrXreference: the average value of X chromosome capture region reads ratio in female's fetal data library；

S.D.%ChrX reference: in female's fetal data library the standard side of X chromosome capture region reads ratio Difference.

Then it makes the following judgment: if X chromosome Z-score >=3 of pregnant woman to be measured and pregnant woman's Y chromosome to be measured capture Region reads ratio < 0.00345, then the X chromosome number increase of pregnant woman fetus to be measured, Y chromosome number are normal；If to Survey X chromosome Z-score >=3 of pregnant woman and ratio >=0.00345 pregnant woman's Y chromosome capture region reads to be measured, then it is to be measured pregnant The Y chromosome number of woman fetus increases, X chromosome number is normal；If the X chromosome Z-score < 3 of -3 < pregnant woman to be measured, Then the X chromosome of pregnant woman fetus to be measured and Y chromosome number are normal；If the X chromosome Z-score of pregnant woman to be measured≤- 3, Then the X chromosome number of pregnant woman fetus to be measured is reduced, and Y chromosome number is normal.

The average value of Y chromosome capture region reads ratio of male fetus of giving a birth out in 400 normal pregnancies is 0.00345。

When sex of foetus is male, X chromosome number increases or Y chromosome number increase will cause chromosome aneuploidy Body disease.X chromosome number increase will cause Klinefelter syndrome.Y chromosome number increase will cause XYY syndrome.

It is non-whole whether the noninvasive antenatal detection fetus to be measured of embodiment 3, the kit prepared using embodiment 1 suffers from chromosome Times body disease

Sample to be tested is that high risk gravida peripheral blood sample (number 7), 2 18- three-bodies are sieved by 1 13- patau syndrome Tang High risk gravida peripheral blood sample (number is 1 and 6) is sieved by syndrome Tang, high risk gravida peripheral blood sample is sieved by 2 21- patau syndrome Tang High risk gravida peripheral blood sample (number 4), 1 gram are sieved by this (number is 2 and 3), 1 poly-x female syndrome (specially XXX) Tang High risk gravida peripheral blood sample (number 5), 1 Turner syndrome sieve (specially in Lan Feierte syndrome (specially XXY) Tang XO) high risk gravida periphery is sieved by Tang's sieve high risk gravida peripheral blood sample (number 9) and 1 XYY syndrome (specially XYY) Tang Blood sample (number 8).

1, the noninvasive antenatal detection of kit prepared using embodiment 1

Sample to be tested is taken, is detected according to the method that embodiment 2 is established.

Average sequencing depth testing result is shown in Table the 2nd column in 3.Sex of foetus is shown in Table the 3rd column in 3, and (F is women, and M is male Property).Y chromosome capture region reads ratio is shown in Table the 4th column in 3.The target chromosome Z-score of pregnant woman is shown in Table the 5th column in 3. For sequencing data amount in 1G, the coverage of 20X or more is shown in Table the 6th column in 3.Testing result is shown in Table the 7th column in 3.

Table 3

2, amniotic fluid gold mark detects

Amniotic fluid gold mark testing result is shown in Table the 8th column in 3.

The result shows that the noninvasive antenatal testing result and amniotic fluid gold mark that are carried out using kit prepared by embodiment 1 are detected As a result completely the same.It can be seen that whether the noninvasive antenatal detection fetus to be measured of the kit that can be prepared using embodiment 1 is suffered from Prenatal diagnosis (such as 13- patau syndrome, E trisomy, 21- patau syndrome, Turner syndrome, gram Lan Feierte syndrome).

<110>Beijing Mai Jinuo Gene science limited liability company

<120>it is a kind of for Non-invasive detection fetal chromosomal to be measured whether be aneuploid kit and its application specific probe group

<160> 508

<170> PatentIn version 3.5

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<213>artificial sequence

<400> 22

gactacatgg gacataataa gcaggagtct tggaggatgg ataactgtag aaagtgggca 60

tgcctgggca aggtacagcc actctgaggg cactacgtcg taggttcc 108

<210> 23

<211> 108

<212> DNA

<213>artificial sequence

<400> 23

gactacatgg gacatagtgg gcatgcctgg gcaaggtaca gccactctga gggcacttca 60

ggtgacataa agtggacagc agcaacttgg gagtacgtcg taggttcc 108

<210> 24

<211> 108

<212> DNA

<213>artificial sequence

<400> 24

gactacatgg gacatagcaa actcagtttt tcacagtgaa atgattcctg taggatcctt 60

tccctttgac agctgcagaa gcgtcagtgc tagtacgtcg taggttcc 108

<210> 25

<211> 108

<212> DNA

<213>artificial sequence

<400> 25

gactacatgg gacatttgtc cttctcattg cctatttctt gggtgaaatg ggtctccatg 60

agcacggatc ccaggagtct gggtctcaga cgatacgtcg taggttcc 108

<210> 26

<211> 108

<212> DNA

<213>artificial sequence

<400> 26

gactacatgg gacataggaa cagcgactgg ccggtttgta gatgtcgaag caccagtttc 60

caggaattga aagcatggct cccgcagttg cgctacgtcg taggttcc 108

<210> 27

<211> 108

<212> DNA

<213>artificial sequence

<400> 27

gactacatgg gacatgactc tgccaccagg ccccggggcc accactcact ggatcaaatc 60

ctttcttctg gaaaatttca attcaagaca ggctacgtcg taggttcc 108

<210> 28

<211> 108

<212> DNA

<213>artificial sequence

<400> 28

gactacatgg gacattatgc cacaagcaac catccttatg tttgtggtgg gaattctaga 60

gggtgattaa ttgtcttagg gaggcagagc aggtacgtcg taggttcc 108

<210> 29

<211> 108

<212> DNA

<213>artificial sequence

<400> 29

gactacatgg gacatgctgg agacagacct ctccttcatc cttcactgca tccatcctct 60

cccttgcctt gtagacaggg gtaccttggc atgtacgtcg taggttcc 108

<210> 30

<211> 108

<212> DNA

<213>artificial sequence

<400> 30

gactacatgg gacatccatc ccaattcttc agtatttgag ttggaaaaaa tctgcacaga 60

tgacacatta tggagagtta gtgagattgg gtgtacgtcg taggttcc 108

<210> 31

<211> 108

<212> DNA

<213>artificial sequence

<400> 31

gactacatgg gacatccgga gagaacccca gaaaagccca ttaaacattt atttaggtaa 60

aatcactgtg tcattaactt atctccatac acatacgtcg taggttcc 108

<210> 32

<211> 108

<212> DNA

<213>artificial sequence

<400> 32

gactacatgg gacataggtg ctgagagctt ccattaggac tgacctctgg aatgggcccc 60

tcaccgggcc tgcgtccacc ccactgctgt ccttacgtcg taggttcc 108

<210> 33

<211> 108

<212> DNA

<213>artificial sequence

<400> 33

gactacatgg gacataggct cgggcggctc ccatggcagg ccagctaggt accctctctg 60

aaccttggtt ccccttctat agaggaggtc atgtacgtcg taggttcc 108

<210> 34

<211> 108

<212> DNA

<213>artificial sequence

<400> 34

gactacatgg gacattcatg agacaagagg agggggaagt gcccagcaca gtgcttggca 60

catggcactg agtgagtgcc agtgtccccc ctttacgtcg taggttcc 108

<210> 35

<211> 108

<212> DNA

<213>artificial sequence

<400> 35

gactacatgg gacatttgga actttgaggc tgctgactcc ctcctgggaa ccccaccttc 60

cagtcatctg cattccctct tgtttcctct ccttacgtcg taggttcc 108

<210> 36

<211> 108

<212> DNA

<213>artificial sequence

<400> 36

gactacatgg gacattgcct tccagcccct tagtgtcctc ctcgtagcag agcacaggga 60

ttggcaagca gtgagcactc agccaacgcc atctacgtcg taggttcc 108

<210> 37

<211> 108

<212> DNA

<213>artificial sequence

<400> 37

gactacatgg gacatggagt taagtgggat tcatctgttt ttttcattta taatgggatg 60

cagtggctta aaaatcctat caccaagcca aattacgtcg taggttcc 108

<210> 38

<211> 108

<212> DNA

<213>artificial sequence

<400> 38

gactacatgg gacattttta aggtttccac agctggattg tttactaggg ccaattgtca 60

ccaatggcag taaaactcat tttctgggga atatacgtcg taggttcc 108

<210> 39

<211> 108

<212> DNA

<213>artificial sequence

<400> 39

gactacatgg gacataatgt gttttgaagg tttgctgcat tggaagttga caataaaatt 60

acagcacgtg cccaccatct atcccccacc ctctacgtcg taggttcc 108

<210> 40

<211> 108

<212> DNA

<213>artificial sequence

<400> 40

gactacatgg gacatgctgg gacttaagcc cagttttgct tatcatgata tctgctgtct 60

ttcctgacac cttaatataa attttgggag gaatacgtcg taggttcc 108

<210> 41

<211> 108

<212> DNA

<213>artificial sequence

<400> 41

gactacatgg gacattaacc ctatcagcca ctggtagcag caggcatcct ccgatgtgcc 60

tatcatctct gatgacaaga gcaaatggtc ttctacgtcg taggttcc 108

<210> 42

<211> 108

<212> DNA

<213>artificial sequence

<400> 42

gactacatgg gacatatcct tcaagaagcc tctccaagcc acacggttta agtcttgaag 60

tccacaattc tctgaatgta ttgacagtct tactacgtcg taggttcc 108

<210> 43

<211> 108

<212> DNA

<213>artificial sequence

<400> 43

gactacatgg gacatcttac cattatttat tccccatgta aaccttccct ttcaaatcag 60

cacagggttt agccaattca aaattcaatt ctatacgtcg taggttcc 108

<210> 44

<211> 108

<212> DNA

<213>artificial sequence

<400> 44

gactacatgg gacattgcgg atgctgcagc ccagcactcc tccaggtttg gaaaaggcgg 60

ctacgctggg caagtttgga aagttctggg agatacgtcg taggttcc 108

<210> 45

<211> 108

<212> DNA

<213>artificial sequence

<400> 45

gactacatgg gacattaggg tgtcttctgg gttcgtatga gtgtgtggat gttaagcggt 60

agtaaaaata gcaactgaag caaaaaaaaa cattacgtcg taggttcc 108

<210> 46

<211> 108

<212> DNA

<213>artificial sequence

<400> 46

gactacatgg gacatatgtt atgtccaaga aaggcattat ccatgcccag atggtaagga 60

gaatttttga aagccagaaa gatagaggac atttacgtcg taggttcc 108

<210> 47

<211> 108

<212> DNA

<213>artificial sequence

<400> 47

gactacatgg gacattttga ccaaggtgac atcttatgtt ctagaataat atcatatggg 60

tcccagaaaa tcaagtttac ccagatttca aaatacgtcg taggttcc 108

<210> 48

<211> 108

<212> DNA

<213>artificial sequence

<400> 48

gactacatgg gacatgatat gcagtctttt gtagactgca aggctctcct cacagcaggc 60

aatgtgcaca aagcaccatc cacaccacgg ttgtacgtcg taggttcc 108

<210> 49

<211> 108

<212> DNA

<213>artificial sequence

<400> 49

gactacatgg gacatacagc aggcaatgtg cacaaagcac catccacacc acggttgaga 60

catagccgcc ctgccttaca ttatgcccat tcatacgtcg taggttcc 108

<210> 50

<211> 108

<212> DNA

<213>artificial sequence

<400> 50

gactacatgg gacatgtccc ctgttttact aatgtctgta gagcagggct tcagatttca 60

tagccaacag cttcaagaac tttcaaaaac gtgtacgtcg taggttcc 108

<210> 51

<211> 108

<212> DNA

<213>artificial sequence

<400> 51

gactacatgg gacatcagat ttcatagcca acagcttcaa gaactttcaa aaacgtggca 60

ggcaggactc ccttgtgggt ttgggttgct ttgtacgtcg taggttcc 108

<210> 52

<211> 108

<212> DNA

<213>artificial sequence

<400> 52

gactacatgg gacataacag gtgactggaa tgaagggatg agctgattca ggcaaggatc 60

aataaaaaac acttgcgcag attcgaagtt acttacgtcg taggttcc 108

<210> 53

<211> 108

<212> DNA

<213>artificial sequence

<400> 53

gactacatgg gacatgcttg aaaacagtgc agcaggtagt caatgccccc ttgttatctt 60

cccgtagaat ggactcgacc aagatctatg atctacgtcg taggttcc 108

<210> 54

<211> 108

<212> DNA

<213>artificial sequence

<400> 54

gactacatgg gacattgatc taagagcttc aaaaattaaa tctaccttgc catctaatta 60

aacatcccca aacactagtg tgttcaaggt gcctacgtcg taggttcc 108

<210> 55

<211> 108

<212> DNA

<213>artificial sequence

<400> 55

gactacatgg gacatccaag cccactataa aaaatgaaca gggttcactt gggagaaaag 60

aaagattcag gaatgggggt ggaaggacac ttctacgtcg taggttcc 108

<210> 56

<211> 108

<212> DNA

<213>artificial sequence

<400> 56

gactacatgg gacatagctg ggggaactgc tgtcccccac cctcatcctt gctagagaag 60

gagatctaat aggatttgta aatctgtagg ccatacgtcg taggttcc 108

<210> 57

<211> 108

<212> DNA

<213>artificial sequence

<400> 57

gactacatgg gacatagacc tccagaatcg ctgcctaaga tagttcatcc aacccttcca 60

gtcatttggc acaagagagt cagttttttg aattacgtcg taggttcc 108

<210> 58

<211> 108

<212> DNA

<213>artificial sequence

<400> 58

gactacatgg gacatagtag gggaggctgg atctgttttg tgatgggacc caaaaccagc 60

ccttagattg aactgtttca ctgacacttg tcatacgtcg taggttcc 108

<210> 59

<211> 108

<212> DNA

<213>artificial sequence

<400> 59

gactacatgg gacatctgta ataaaatctt gccagatgtg attctctgca tttttgtcaa 60

tgggacaact ggactgcaga tattgagtag ttctacgtcg taggttcc 108

<210> 60

<211> 108

<212> DNA

<213>artificial sequence

<400> 60

gactacatgg gacatagttc aaggtcacat cacgacagga atgggaccat cccaccagaa 60

gctatctgac ccctagcaca atgttgttcc aaatacgtcg taggttcc 108

<210> 61

<211> 108

<212> DNA

<213>artificial sequence

<400> 61

gactacatgg gacatcttta cccacaacct ttccccacta gctgaacttc gtgattttgg 60

ggatgctaga ggacaatctg accaaaactt gactacgtcg taggttcc 108

<210> 62

<211> 108

<212> DNA

<213>artificial sequence

<400> 62

gactacatgg gacattaaac gtttacgaag agtagatttt tttgctaaga cttcttgaaa 60

agtctggcgg gtttcttttt tttttttttt ctttacgtcg taggttcc 108

<210> 63

<211> 108

<212> DNA

<213>artificial sequence

<400> 63

gactacatgg gacatgcatg attctgcagc tgctcgtctc tcagtgtctc ctgtgctgct 60

tcaacttgaa gttttccttc atagcacctt ttttacgtcg taggttcc 108

<210> 64

<211> 108

<212> DNA

<213>artificial sequence

<400> 64

gactacatgg gacatttata tttttggcaa aatgagactc tgtcttcata gataaatcca 60

gctacctaca gttgtcagtc acgtaagtga tattacgtcg taggttcc 108

<210> 65

<211> 108

<212> DNA

<213>artificial sequence

<400> 65

gactacatgg gacatcatca cccaatggac gattcagaat ctccatctcc ccacctttct 60

tcgcataaca tggccctcta attcaatgtt ctctacgtcg taggttcc 108

<210> 66

<211> 108

<212> DNA

<213>artificial sequence

<400> 66

gactacatgg gacataacct tctcctatat ctttctttcc cattcccttt cctcatacct 60

acctccacct tgttggctca gtaatcatct ctgtacgtcg taggttcc 108

<210> 67

<211> 108

<212> DNA

<213>artificial sequence

<400> 67

gactacatgg gacatagtac taaaacagtg tctacctaag tctcctgcct tacattggag 60

tgaaataatt aattacacag tggagtgtct ggttacgtcg taggttcc 108

<210> 68

<211> 108

<212> DNA

<213>artificial sequence

<400> 68

gactacatgg gacatgcagg agggcaggct agtataatat gaagtagatg tggaagagta 60

gaaaacagag agagcctagg cttgtttaca cggtacgtcg taggttcc 108

<210> 69

<211> 108

<212> DNA

<213>artificial sequence

<400> 69

gactacatgg gacatcacgg agggggtcag tctcaaaaag aagaaaacag gatgggatgt 60

acccaataat tcactcgaaa cagtaaacag ccatacgtcg taggttcc 108

<210> 70

<211> 108

<212> DNA

<213>artificial sequence

<400> 70

gactacatgg gacattttct tagactttta acaggtacaa atcctcagca gccatgttgt 60

ttatctaaat tcagcccaga gtctgtgtca tggtacgtcg taggttcc 108

<210> 71

<211> 108

<212> DNA

<213>artificial sequence

<400> 71

gactacatgg gacattcggt cagatggggg cgctcaagag cccaggagga aagtcaagct 60

tctttatagg cagaaagtaa tatggtacct gaatacgtcg taggttcc 108

<210> 72

<211> 108

<212> DNA

<213>artificial sequence

<400> 72

gactacatgg gacatccatg ctccagagac aggccttcag accccgagag gtacttgaac 60

acacaacagc aagttcaagt tcacattcaa gtgtacgtcg taggttcc 108

<210> 73

<211> 108

<212> DNA

<213>artificial sequence

<400> 73

gactacatgg gacatgggat cttcttaaac caataaaaat aaacattttg tgatctaaga 60

cttcatagat gctaacattg gaacggagaa gattacgtcg taggttcc 108

<210> 74

<211> 108

<212> DNA

<213>artificial sequence

<400> 74

gactacatgg gacatgtttt ctctgttatt cacccagttt gggaaggagt gccctgtgct 60

ctccccaacc ccaggctcac ttccctaccc tcgtacgtcg taggttcc 108

<210> 75

<211> 108

<212> DNA

<213>artificial sequence

<400> 75

gactacatgg gacatcacag cttaggcaca aaagcccagc ttcccatctg ctgtgtgggt 60

gggtggctcc tctgcaggtc aaagcgtgat cattacgtcg taggttcc 108

<210> 76

<211> 108

<212> DNA

<213>artificial sequence

<400> 76

gactacatgg gacatatctt ctcttttttc cttactccca ctttgcacct attacctagc 60

tttctctata aagaatttat atcccactct tcttacgtcg taggttcc 108

<210> 77

<211> 108

<212> DNA

<213>artificial sequence

<400> 77

gactacatgg gacatttacc tagctttctc tataaagaat ttatatccca ctcttctttg 60

ttcaagctgg cctttctggt gagggtgata tgatacgtcg taggttcc 108

<210> 78

<211> 108

<212> DNA

<213>artificial sequence

<400> 78

gactacatgg gacattactt cagagagaga aaaatagctt gttcatggag ttcagagttg 60

cctgatccct caggagccct gacatttttt taatacgtcg taggttcc 108

<210> 79

<211> 108

<212> DNA

<213>artificial sequence

<400> 79

gactacatgg gacatggcaa gtccaaagct ctgagggaga gaaagagagg agcaggagag 60

aatggagagg gagagactat aaatttggaa aattacgtcg taggttcc 108

<210> 80

<211> 108

<212> DNA

<213>artificial sequence

<400> 80

gactacatgg gacatggaaa gaacttattc cacacttagc ccaatgttta agacaactta 60

ttcgaaatgg gactatagaa ttattagaaa tggtacgtcg taggttcc 108

<210> 81

<211> 108

<212> DNA

<213>artificial sequence

<400> 81

gactacatgg gacatcacta aaaagagcca ttgctaagtg attttactgc cgcttagaaa 60

tttcaatctc aagggctggg gttcccagag cggtacgtcg taggttcc 108

<210> 82

<211> 108

<212> DNA

<213>artificial sequence

<400> 82

gactacatgg gacatagcgg gagattatgt caaactgcaa ctcatggtgc cagcacccct 60

tgacgaacac aaaccactgt tttgtacttc agatacgtcg taggttcc 108

<210> 83

<211> 108

<212> DNA

<213>artificial sequence

<400> 83

gactacatgg gacatggtac cctctgtggc tgccttctgc tcccatgcca tgccattgac 60

catttaccct atattcctct tcccagactt tagtacgtcg taggttcc 108

<210> 84

<211> 108

<212> DNA

<213>artificial sequence

<400> 84

gactacatgg gacatccatt gaccatttac cctatattcc tcttcccaga ctttagattt 60

gaatttcagg aaaagaaaat tttcaaggaa aaatacgtcg taggttcc 108

<210> 85

<211> 108

<212> DNA

<213>artificial sequence

<400> 85

gactacatgg gacatgataa gttgttctct gggagataac agtagatctg ccagaaaact 60

cttcaccaat ggcccaaagt gctgacttca tggtacgtcg taggttcc 108

<210> 86

<211> 108

<212> DNA

<213>artificial sequence

<400> 86

gactacatgg gacatagaaa actcttcacc aatggcccaa agtgctgact tcatggttaa 60

gtggcatgtg gaaataaatg gcattcagaa agctacgtcg taggttcc 108

<210> 87

<211> 108

<212> DNA

<213>artificial sequence

<400> 87

gactacatgg gacattttgg ccaagacaga tggcttgaag tgaggagcca tgagagtagc 60

aatcagaaag tcaagaaccc cgatgccaat ttatacgtcg taggttcc 108

<210> 88

<211> 108

<212> DNA

<213>artificial sequence

<400> 88

gactacatgg gacatgaaaa gtagcctctg gtcaggggtc atatattttt ttcagccaaa 60

cagatcctct tgggctccag tggcctcctc ccctacgtcg taggttcc 108

<210> 89

<211> 108

<212> DNA

<213>artificial sequence

<400> 89

gactacatgg gacatcagcc aaacagatcc tcttgggctc cagtggcctc ctccccaact 60

ttacctgtga gatcagggat tatagcaaaa ggatacgtcg taggttcc 108

<210> 90

<211> 108

<212> DNA

<213>artificial sequence

<400> 90

gactacatgg gacatttagt ggctttcttt cctaaacctg cttctctctt cagttgcctg 60

gctggtttgg ggttgctcct ttagtatcac gcttacgtcg taggttcc 108

<210> 91

<211> 108

<212> DNA

<213>artificial sequence

<400> 91

gactacatgg gacatacgct caagtgggac attttagaca gctggggctt ccaggttcaa 60

ccagctcctc tcttccatta actcaggaac acctacgtcg taggttcc 108

<210> 92

<211> 108

<212> DNA

<213>artificial sequence

<400> 92

gactacatgg gacatgcatt ccttgcatgg gaaccttgac cttctgctcc tcataaagca 60

accacaaagg cttccttttt caggcagatt cggtacgtcg taggttcc 108

<210> 93

<211> 108

<212> DNA

<213>artificial sequence

<400> 93

gactacatgg gacatcataa agcaaccaca aaggcttcct ttttcaggca gattcggact 60

gcaaaatttt tcaccaagtg catgttctca aattacgtcg taggttcc 108

<210> 94

<211> 108

<212> DNA

<213>artificial sequence

<400> 94

gactacatgg gacatcccaa gtgtcagggt gaaattggga tggaaactgg gtatgaagta 60

tggaatgaag tatgtctaag accttgacat gaatacgtcg taggttcc 108

<210> 95

<211> 108

<212> DNA

<213>artificial sequence

<400> 95

gactacatgg gacatatgaa gtatggaatg aagtatgtct aagaccttga catgaaggaa 60

ggggctggca aggagcatga gataagggag ggttacgtcg taggttcc 108

<210> 96

<211> 108

<212> DNA

<213>artificial sequence

<400> 96

gactacatgg gacatttatg gcaacatgat gaagatatcc atctccagcc agtactacag 60

cagcctctgg tgttgattct ggacttgggg gtatacgtcg taggttcc 108

<210> 97

<211> 108

<212> DNA

<213>artificial sequence

<400> 97

gactacatgg gacatgggaa tatgtgagtc tgcttaggaa ggagtaaagt ggctgtggat 60

tcctgctctt attgtcgaag cgaggccagt acatacgtcg taggttcc 108

<210> 98

<211> 108

<212> DNA

<213>artificial sequence

<400> 98

gactacatgg gacataaaag aaacaaaaca tttcaccttt ttccctgtag aagccagagt 60

gtgcttctca caaaagcctg tgcaacctcc gattacgtcg taggttcc 108

<210> 99

<211> 108

<212> DNA

<213>artificial sequence

<400> 99

gactacatgg gacatgtata aatcctagag acagtaatct gccccagcaa acacagactt 60

ctgtggaaca agctgctgta tgtaggaacc aactacgtcg taggttcc 108

<210> 100

<211> 108

<212> DNA

<213>artificial sequence

<400> 100

gactacatgg gacatgctta tcatcaaatt gataatcaca ttcacctaat aaacccaacg 60

gatgagacac tgtttcctgg aataaatagc aaatacgtcg taggttcc 108

<210> 101

<211> 108

<212> DNA

<213>artificial sequence

<400> 101

gactacatgg gacattttta aaaatgcaag agaagtgtac ttgctggata aagtacaaga 60

ttttggcttc acatctgtag gtccaggggc gtgtacgtcg taggttcc 108

<210> 102

<211> 108

<212> DNA

<213>artificial sequence

<400> 102

gactacatgg gacatagatt atacagcttc attctctaac acttaaagtc cgatagttgg 60

gcttgattgc ccacattagg tctggttccc gagtacgtcg taggttcc 108

<210> 103

<211> 108

<212> DNA

<213>artificial sequence

<400> 103

gactacatgg gacatcagag gccactgcaa tgccttgact acctacagtg attattctct 60

ggccttaaac cgctgaagtg aagctaccaa gtgtacgtcg taggttcc 108

<210> 104

<211> 108

<212> DNA

<213>artificial sequence

<400> 104

gactacatgg gacataccca tacggagcgg ccaggaggaa ccatttctga catccaccaa 60

atccctcaca ttggcagtgg cctattctac ttatacgtcg taggttcc 108

<210> 105

<211> 108

<212> DNA

<213>artificial sequence

<400> 105

gactacatgg gacatctaga agagaagagg ccctctccag aataccatgt cctctctctt 60

ccaaatttgg ggtcagtcct gtttatactt cagtacgtcg taggttcc 108

<210> 106

<211> 108

<212> DNA

<213>artificial sequence

<400> 106

gactacatgg gacattggga tcaacccaag acagattttg atgaggagtt accattaaca 60

aaatcatcta aatcaggatg gcaggctcaa gtgtacgtcg taggttcc 108

<210> 107

<211> 108

<212> DNA

<213>artificial sequence

<400> 107

gactacatgg gacatgtaca tttctctttg atattcagta gtcagcacat acccaatcac 60

aacaccgctg ctttctttgc accctcctgt tggtacgtcg taggttcc 108

<210> 108

<211> 108

<212> DNA

<213>artificial sequence

<400> 108

gactacatgg gacatcagta ggccagatca gacagatttg actttttcct acgtgcacta 60

agaaagcaat ggagccctcc aacccccacc ccctacgtcg taggttcc 108

<210> 109

<211> 108

<212> DNA

<213>artificial sequence

<400> 109

gactacatgg gacatggcat aaaattgatg ggaagagaat atgtagagca atcttgaggt 60

gctattaaat agttactggt ctatttcttt ctttacgtcg taggttcc 108

<210> 110

<211> 108

<212> DNA

<213>artificial sequence

<400> 110

gactacatgg gacatgcttg tacaaatctg aagtatgtga atttgggcag catctcccct 60

tgataagctg ttaatcaagg attattattt ggctacgtcg taggttcc 108

<210> 111

<211> 108

<212> DNA

<213>artificial sequence

<400> 111

gactacatgg gacattggtc atttagggaa agccagttgg accttctgag gcaattcaat 60

tctgttcttt tcaggacagc tctatggagt cgatacgtcg taggttcc 108

<210> 112

<211> 108

<212> DNA

<213>artificial sequence

<400> 112

gactacatgg gacatagcag ttcagggcgc tctgcctgat gcagaccacc tactcgggga 60

ccagactgga acgtggcttt ggggatgttc aggtacgtcg taggttcc 108

<210> 113

<211> 108

<212> DNA

<213>artificial sequence

<400> 113

gactacatgg gacataaaca gaggaggagt caacacagaa aactcatggt actagaaatg 60

aacacaacag gacttcagaa aatcgtgcct acttacgtcg taggttcc 108

<210> 114

<211> 108

<212> DNA

<213>artificial sequence

<400> 114

gactacatgg gacatgtagg tagatacaga ccctgggata gcagtgcctg ggggtctctc 60

tgggcagaga tgtcgggtca aatggacatg acctacgtcg taggttcc 108

<210> 115

<211> 108

<212> DNA

<213>artificial sequence

<400> 115

gactacatgg gacatcccct ggctctggcc ctgcctagca cagtccctgt cccatgctct 60

gacaagttct tgagggagcc cagtgactca tggtacgtcg taggttcc 108

<210> 116

<211> 108

<212> DNA

<213>artificial sequence

<400> 116

gactacatgg gacataaaaa aatacaatct tttttttgca agttaaaact gagctaatga 60

cactgggtgg ggctcagtgt tgaggactcc ccatacgtcg taggttcc 108

<210> 117

<211> 108

<212> DNA

<213>artificial sequence

<400> 117

gactacatgg gacatgtgat gtgagcacta tgcaggttca gtagaggaag gagtggggtg 60

acatgttatg gaggttcagt aggggaagga atgtacgtcg taggttcc 108

<210> 118

<211> 108

<212> DNA

<213>artificial sequence

<400> 118

gactacatgg gacattttcc ttacagtggg aggttatatg taggtgggat atatcaaagt 60

tactgagcac caaattttat ttttctaagt aagtacgtcg taggttcc 108

<210> 119

<211> 108

<212> DNA

<213>artificial sequence

<400> 119

gactacatgg gacatctaaa aacccagctt aaatgacagg agaggcggca gaccgtaaaa 60

ccttattcag tgacctaaga gtggcctctg cgttacgtcg taggttcc 108

<210> 120

<211> 108

<212> DNA

<213>artificial sequence

<400> 120

gactacatgg gacattacaa acttcaaaat tctgatgatc gccaacatgg tgaagcagga 60

tccactgggc ttataggccc atctgaaaac gggtacgtcg taggttcc 108

<210> 121

<211> 108

<212> DNA

<213>artificial sequence

<400> 121

gactacatgg gacatactga cataagaggc acagcatctg atggtcttat caaatcaatc 60

catcttctgt ctcttccagg ggagtgctac gcatacgtcg taggttcc 108

<210> 122

<211> 108

<212> DNA

<213>artificial sequence

<400> 122

gactacatgg gacatgtgct cagtccctcc acctgaattc tcacactgag ctcattattt 60

cctctgacac cttcccactg atccctatcc gcctacgtcg taggttcc 108

<210> 123

<211> 108

<212> DNA

<213>artificial sequence

<400> 123

gactacatgg gacatatcca ctggttccca gcgggaacaa catgacctgt gctccatgac 60

cttgctcagc tatcagtagg gccgctctac gtgtacgtcg taggttcc 108

<210> 124

<211> 108

<212> DNA

<213>artificial sequence

<400> 124

gactacatgg gacattgctc tgcagagccc aggaaaataa taaaaccttt atgaccaaga 60

gtactatttc atttcctttc taaacgaaac ccatacgtcg taggttcc 108

<210> 125

<211> 108

<212> DNA

<213>artificial sequence

<400> 125

gactacatgg gacatactct tgaaaacact gttgaattga catggagtgt ctgatttgat 60

ggaagccccg tagacctgga acggtatctg ggatacgtcg taggttcc 108

<210> 126

<211> 108

<212> DNA

<213>artificial sequence

<400> 126

gactacatgg gacattctct ggcaaaaaga aacaaaactc caaaaacttg aaaaagccaa 60

tatcctgcct agttgtatat aagggaacag gaatacgtcg taggttcc 108

<210> 127

<211> 108

<212> DNA

<213>artificial sequence

<400> 127

gactacatgg gacattaggg tgggggtagt ttgggtggtc ttcctgcaag aggttacatg 60

gttcatcatc attaggtttt aatggaaata cgttacgtcg taggttcc 108

<210> 128

<211> 108

<212> DNA

<213>artificial sequence

<400> 128

gactacatgg gacattggtc aggaattccc ggtgcagcag aatggagcca agattctgtg 60

cagaccaccc ttttcttttc ttcatcaggc acctacgtcg taggttcc 108

<210> 129

<211> 108

<212> DNA

<213>artificial sequence

<400> 129

gactacatgg gacataaatg ctacagggaa ttacgcatgt acggtccatc agtgttaatg 60

ggtgaataaa ccttacagtt tgttaagtaa tgttacgtcg taggttcc 108

<210> 130

<211> 108

<212> DNA

<213>artificial sequence

<400> 130

gactacatgg gacattccta actgctctac aaatcagcac tcctgtgtat agcttatgcc 60

cataaaacta gaacaacaat aaaaacatta gcatacgtcg taggttcc 108

<210> 131

<211> 108

<212> DNA

<213>artificial sequence

<400> 131

gactacatgg gacatttcat ctctcccctc tcaacgaagg cataatgcta agggcgctgg 60

agttcccatt gacaagataa ccacgtcctc tgttacgtcg taggttcc 108

<210> 132

<211> 108

<212> DNA

<213>artificial sequence

<400> 132

gactacatgg gacatggaga aatgcatagc atgctttatg tctgagtgaa tcttttgaaa 60

cattgttttc aaaaagtatt cacgctcttc acttacgtcg taggttcc 108

<210> 133

<211> 108

<212> DNA

<213>artificial sequence

<400> 133

gactacatgg gacatggcaa gagcatacaa atacaaggcc atatggtgtc cttttctgag 60

gtaagaggca ataacccaac acacttagga gtttacgtcg taggttcc 108

<210> 134

<211> 108

<212> DNA

<213>artificial sequence

<400> 134

gactacatgg gacatgtaac tcaaatatca gctgcacccc agccttcgcg gaccgcactg 60

ttttaacctt gtgctaactt gtgtagagat cagtacgtcg taggttcc 108

<210> 135

<211> 108

<212> DNA

<213>artificial sequence

<400> 135

gactacatgg gacatacctg gcattgcagg gctctcaatg gagacagctg atgtcgccct 60

gctcctcctg agcctcctcc atcgtgaaaa tattacgtcg taggttcc 108

<210> 136

<211> 108

<212> DNA

<213>artificial sequence

<400> 136

gactacatgg gacatttatt ctctggcctt aaaccgctga agtgaagcta ccaagtgggc 60

agacagcatt ttgctttaga gtttcaggac ttttacgtcg taggttcc 108

<210> 137

<211> 108

<212> DNA

<213>artificial sequence

<400> 137

gactacatgg gacatttgta ttgtctgaga ttgtatgcag cctcagagca catgtccttt 60

tcctcctcca catttactac aacattataa aaatacgtcg taggttcc 108

<210> 138

<211> 108

<212> DNA

<213>artificial sequence

<400> 138

gactacatgg gacattccct tctgtaattt cttaaggaca tgggaattta tccattgaag 60

tgtttcaatt ctcttccaag tggccaagga gaatacgtcg taggttcc 108

<210> 139

<211> 108

<212> DNA

<213>artificial sequence

<400> 139

gactacatgg gacatgaaag actggacacc tgccatctta gcacctgaaa gctacgaggc 60

aggttgaaag aatcctgagc aatgcgtcgg agatacgtcg taggttcc 108

<210> 140

<211> 108

<212> DNA

<213>artificial sequence

<400> 140

gactacatgg gacatctctc tcttccaaat ttggggtcag tcctgtttat acttcagtat 60

actgaagctg agttgccatt tcagccttgt ctttacgtcg taggttcc 108

<210> 141

<211> 108

<212> DNA

<213>artificial sequence

<400> 141

gactacatgg gacatccata aatctatgta accacttcct tttacattca ttatgtggga 60

gcagaagcaa ctctcagaat cagaagcagc atctacgtcg taggttcc 108

<210> 142

<211> 108

<212> DNA

<213>artificial sequence

<400> 142

gactacatgg gacattacca acatcagaca gacacactgg gttcactaat gcataatttt 60

tcaaactatc tatggagaca gctgtcgttt tcttacgtcg taggttcc 108

<210> 143

<211> 108

<212> DNA

<213>artificial sequence

<400> 143

gactacatgg gacatttggg ctaatctttc gcaagccagt aaatgtgcat ttatctgttt 60

agacatatat ttatgtgcac gcatgcacta agttacgtcg taggttcc 108

<210> 144

<211> 108

<212> DNA

<213>artificial sequence

<400> 144

gactacatgg gacatactgt gtatctaagg gtttgatttt gtttgatcta tgtgagtttt 60

tactgctaaa tatttgaaag gaggagaatt ttttacgtcg taggttcc 108

<210> 145

<211> 108

<212> DNA

<213>artificial sequence

<400> 145

gactacatgg gacatagtca ttcattcctt cctcagtgag ttctgtccct ggccctaatt 60

aaaattaatg tcctgactag gaagtgacta ccctacgtcg taggttcc 108

<210> 146

<211> 108

<212> DNA

<213>artificial sequence

<400> 146

gactacatgg gacattctgc tactctctct ggcccatgct cgggacagaa tagatgctga 60

aagcatctta ggtacagtct cccctggaag gcatacgtcg taggttcc 108

<210> 147

<211> 108

<212> DNA

<213>artificial sequence

<400> 147

gactacatgg gacatatctc cccttgataa gctgttaatc aaggattatt atttggccca 60

attgctccgc aataggtttg acagaggcct cagtacgtcg taggttcc 108

<210> 148

<211> 108

<212> DNA

<213>artificial sequence

<400> 148

gactacatgg gacatcaatt caattctgtt cttttcagga cagctctatg gagtcgagtc 60

ttaatcagtt gcacaaggca ccatgaagtg ctttacgtcg taggttcc 108

<210> 149

<211> 108

<212> DNA

<213>artificial sequence

<400> 149

gactacatgg gacatactcg gggaccagac tggaacgtgg ctttggggat gttcaggagc 60

tggagaacat ggggccatgg gggcactgtg gcttacgtcg taggttcc 108

<210> 150

<211> 108

<212> DNA

<213>artificial sequence

<400> 150

gactacatgg gacataagat agaaagtact ttcctcttag gaaattaaac tatcgaggag 60

agctagggtt gttcttattg tggcatttgg gagtacgtcg taggttcc 108

<210> 151

<211> 108

<212> DNA

<213>artificial sequence

<400> 151

gactacatgg gacatctaga aatgaacaca acaggacttc agaaaatcgt gcctactgca 60

agagtggaaa ggcttcttgg atgtggcaaa acttacgtcg taggttcc 108

<210> 152

<211> 108

<212> DNA

<213>artificial sequence

<400> 152

gactacatgg gacatcccca ttctcagctc ttaggcagaa atcttactgc tcccatctcc 60

ctgatagaag tgtccctggc attggtattt ctgtacgtcg taggttcc 108

<210> 153

<211> 108

<212> DNA

<213>artificial sequence

<400> 153

gactacatgg gacatacagg ctataaacca ccttaacagg cataatgact ttcatgcctc 60

cacctgaagg ggctcattgc aataaggtaa cagtacgtcg taggttcc 108

<210> 154

<211> 108

<212> DNA

<213>artificial sequence

<400> 154

gactacatgg gacatgctgt tttcaagtag aaataaatct gagatatttg aagatatttg 60

gaggcggcaa cagtgagaag agcagaagct gcttacgtcg taggttcc 108

<210> 155

<211> 108

<212> DNA

<213>artificial sequence

<400> 155

gactacatgg gacatattcc acacacctcc tggtggtctt gacaggaaaa ctgatatcag 60

ggggttgttt tcataaacac tgagcattgg gtatacgtcg taggttcc 108

<210> 156

<211> 108

<212> DNA

<213>artificial sequence

<400> 156

gactacatgg gacatccatg ctctgacaag ttcttgaggg agcccagtga ctcatggcat 60

tcgattttcc acacatcttc ttccccccgt ttctacgtcg taggttcc 108

<210> 157

<211> 108

<212> DNA

<213>artificial sequence

<400> 157

gactacatgg gacatagcta atgacactgg gtggggctca gtgttgagga ctccccagcc 60

aaaccaaaat tagcttccag ccaagagctg ctttacgtcg taggttcc 108

<210> 158

<211> 108

<212> DNA

<213>artificial sequence

<400> 158

gactacatgg gacatgctca gagctggggg cccaaagccc agacagcagc tgccacacgt 60

gtagagaaat tccgtcttgg gttggaggcc ccttacgtcg taggttcc 108

<210> 159

<211> 108

<212> DNA

<213>artificial sequence

<400> 159

gactacatgg gacatagtgg ggtgacatgt tatggaggtt cagtagggga aggaatgggg 60

tgatgtgagc actatggagg ttcagtagag gaatacgtcg taggttcc 108

<210> 160

<211> 108

<212> DNA

<213>artificial sequence

<400> 160

gactacatgg gacattgctg aggcactcat accagaggca ctagactgtg tgagtggagt 60

ctgcagcatg gctcctgtcc tggtagctgg acatacgtcg taggttcc 108

<210> 161

<211> 108

<212> DNA

<213>artificial sequence

<400> 161

gactacatgg gacatcagaa ccagtactat tcttttcaag tcatggcttt ccccgttttt 60

ttttggtctg cagctattct gcagggctaa ggatacgtcg taggttcc 108

<210> 162

<211> 108

<212> DNA

<213>artificial sequence

<400> 162

gactacatgg gacatatctc agtgctgact gcacacggga cgctgtccta agcacgggaa 60

cagatacaat ggcatgctta atctgtccct ggctacgtcg taggttcc 108

<210> 163

<211> 108

<212> DNA

<213>artificial sequence

<400> 163

gactacatgg gacatgtgaa gaaaaggcct ttgcccctct cactactgac tccaggttga 60

aactcgaggg gtcccgggcg gctgcagcgc tgctacgtcg taggttcc 108

<210> 164

<211> 108

<212> DNA

<213>artificial sequence

<400> 164

gactacatgg gacatccctg gcttctcctc tcagaggagc tgaatatcaa gctgtcaagt 60

ggggcagcct tgttccccaa gcacacctgc ccttacgtcg taggttcc 108

<210> 165

<211> 108

<212> DNA

<213>artificial sequence

<400> 165

gactacatgg gacattaaca gctggggtta aagatgagag gtgaatgcca acgcgctgtc 60

catagtggcc ttagaaatga cagctgcgat taatacgtcg taggttcc 108

<210> 166

<211> 108

<212> DNA

<213>artificial sequence

<400> 166

gactacatgg gacattcatt atttcctctg acaccttccc actgatccct atccgccgcg 60

ccctccccag ggtcttcttt cgtttgtgcc ttctacgtcg taggttcc 108

<210> 167

<211> 108

<212> DNA

<213>artificial sequence

<400> 167

gactacatgg gacatctcca tgaccttgct cagctatcag tagggccgct ctacgtggac 60

agctgccctt tcaaaagccc agggaactca gaatacgtcg taggttcc 108

<210> 168

<211> 108

<212> DNA

<213>artificial sequence

<400> 168

gactacatgg gacattgatt tgatggaagc cccgtagacc tggaacggta tctgggatat 60

gttactgagt gtgaaaatgc ctatatatgt gtatacgtcg taggttcc 108

<210> 169

<211> 108

<212> DNA

<213>artificial sequence

<400> 169

gactacatgg gacataaaag ccaatatcct gcctagttgt atataaggga acaggaatac 60

gctggcgttc aggaaacgga tgacttgtaa gcttacgtcg taggttcc 108

<210> 170

<211> 108

<212> DNA

<213>artificial sequence

<400> 170

gactacatgg gacatgattc tgtgcagacc acccttttct tttcttcatc aggcaccaca 60

aaaccaactg catcttgttc tgtggcaggt tcttacgtcg taggttcc 108

<210> 171

<211> 108

<212> DNA

<213>artificial sequence

<400> 171

gactacatgg gacatgtgtt aatgggtgaa taaaccttac agtttgttaa gtaatgtaac 60

atctattagg gaagggtgaa ttggttaatg tattacgtcg taggttcc 108

<210> 172

<211> 108

<212> DNA

<213>artificial sequence

<400> 172

gactacatgg gacattttct tttttctgac cttatgccta ttgctttgct aacttgattc 60

ttgttattgg ttttgccaga ctccgtgtct ttgtacgtcg taggttcc 108

<210> 173

<211> 108

<212> DNA

<213>artificial sequence

<400> 173

gactacatgg gacatccgag actggcagaa gtccacagag atcccctggc tcaagaagag 60

atttcttgac tcagaaaagt ggcgggtttg ttctacgtcg taggttcc 108

<210> 174

<211> 108

<212> DNA

<213>artificial sequence

<400> 174

gactacatgg gacatggatg cccacctagg gatgtgatac tttggagcct tccacattac 60

acaacttcgt gtattttatg caggatagaa gaatacgtcg taggttcc 108

<210> 175

<211> 108

<212> DNA

<213>artificial sequence

<400> 175

gactacatgg gacatgcatc tataccatgg agttggttgt ctcttggaga aaatcttccc 60

ttccttagct gttaaaattg ttaaagatag tggtacgtcg taggttcc 108

<210> 176

<211> 108

<212> DNA

<213>artificial sequence

<400> 176

gactacatgg gacattgtcc ttttcctcct ccacatttac tacaacatta taaaaacctt 60

gggccctgta ccctccattc agaagttcgt ctgtacgtcg taggttcc 108

<210> 177

<211> 108

<212> DNA

<213>artificial sequence

<400> 177

gactacatgg gacatttcag tatactgaag ctgagttgcc atttcagcct tgtctttaat 60

tcacaaagaa atggcttggc taccttgaca aagtacgtcg taggttcc 108

<210> 178

<211> 108

<212> DNA

<213>artificial sequence

<400> 178

gactacatgg gacatataat ttttcaaact atctatggag acagctgtcg ttttcttctt 60

ccaactgtca tacctggata cacagtccta atgtacgtcg taggttcc 108

<210> 179

<211> 108

<212> DNA

<213>artificial sequence

<400> 179

gactacatgg gacatgttgg tgtctccagc acaagtactt actcaatttt tgttaagatt 60

acttctaaca gccttgctaa ctggtccttc aggtacgtcg taggttcc 108

<210> 180

<211> 108

<212> DNA

<213>artificial sequence

<400> 180

gactacatgg gacatccccc caccccaacc ccttggggag aagagaatga aggaatgtgc 60

agagagatgc tttaggacat catactgcgt ccatacgtcg taggttcc 108

<210> 181

<211> 108

<212> DNA

<213>artificial sequence

<400> 181

gactacatgg gacatttctt tttttttttt ttttctagca ctgtttgctg cctacctggg 60

tgctgccctg gctaacatgt tatctgtggt gagtacgtcg taggttcc 108

<210> 182

<211> 108

<212> DNA

<213>artificial sequence

<400> 182

gactacatgg gacatgatgc tgaaagcatc ttaggtacag tctcccctgg aaggcaggaa 60

gcaggtgtgc agagcctcca ggcctgtgga ggatacgtcg taggttcc 108

<210> 183

<211> 108

<212> DNA

<213>artificial sequence

<400> 183

gactacatgg gacatgaacc actgttctca aatgtcttga ctggaccaga attcccaata 60

gtgggatgcc ttctgcacag aacttctcaa tgctacgtcg taggttcc 108

<210> 184

<211> 108

<212> DNA

<213>artificial sequence

<400> 184

gactacatgg gacatgggtg tcatctcctt catgggcctc cagggcatta agcttgatcc 60

ctggctcttg gactggcatg gctaatgccc agctacgtcg taggttcc 108

<210> 185

<211> 108

<212> DNA

<213>artificial sequence

<400> 185

gactacatgg gacataccag ggtggtcagc tgctggctgc aggataatgg cagcatatct 60

ggaccattag ctaagcatct caaggctctg aagtacgtcg taggttcc 108

<210> 186

<211> 108

<212> DNA

<213>artificial sequence

<400> 186

gactacatgg gacatctact gcaagagtgg aaaggcttct tggatgtggc aaaactttag 60

ctgtgctcca gataagaagg aggacttgga tgatacgtcg taggttcc 108

<210> 187

<211> 108

<212> DNA

<213>artificial sequence

<400> 187

gactacatgg gacatctact ttaaacacag ttttctgctc cttgacagtc tttcagatac 60

gtttggggaa atcacaaact agtaagggga tgatacgtcg taggttcc 108

<210> 188

<211> 108

<212> DNA

<213>artificial sequence

<400> 188

gactacatgg gacatcatgc ctccacctga aggggctcat tgcaataagg taacagctct 60

tgcagatgcc tctaacttta actatgggaa cactacgtcg taggttcc 108

<210> 189

<211> 108

<212> DNA

<213>artificial sequence

<400> 189

gactacatgg gacatcaagg gtaagttctg ccttcagaca caccagtggc agccaatatt 60

ggatcccatt tctgccacaa ttccagttgt tggtacgtcg taggttcc 108

<210> 190

<211> 108

<212> DNA

<213>artificial sequence

<400> 190

gactacatgg gacatgatat cagggggttg ttttcataaa cactgagcat tgggtaaacc 60

agcttaaggt ggctgaggta taggcagctg ggctacgtcg taggttcc 108

<210> 191

<211> 108

<212> DNA

<213>artificial sequence

<400> 191

gactacatgg gacatcccca gccaaaccaa aattagcttc cagccaagag ctgcttggcc 60

ttaccatttg gggaaggaag ctgaaaaagt cattacgtcg taggttcc 108

<210> 192

<211> 108

<212> DNA

<213>artificial sequence

<400> 192

gactacatgg gacatgaatg gggtgatgtg agcactatgg aggttcagta gaggaaggag 60

tggggtgatg tgttatggag attcagtagg ggatacgtcg taggttcc 108

<210> 193

<211> 108

<212> DNA

<213>artificial sequence

<400> 193

gactacatgg gacatccgtt tttttttggt ctgcagctat tctgcagggc taaggattgc 60

tagtcattag gtaatcatgg aactgagaac cactacgtcg taggttcc 108

<210> 194

<211> 108

<212> DNA

<213>artificial sequence

<400> 194

gactacatgg gacatcacgg gaacagatac aatggcatgc ttaatctgtc cctggccagt 60

cgaactaagg agacacatct gtacattcac atgtacgtcg taggttcc 108

<210> 195

<211> 108

<212> DNA

<213>artificial sequence

<400> 195

gactacatgg gacatgcgct gtccatagtg gccttagaaa tgacagctgc gattaaagat 60

aagaggtgaa cgccaacgtg ctgtccatag tggtacgtcg taggttcc 108

<210> 196

<211> 108

<212> DNA

<213>artificial sequence

<400> 196

gactacatgg gacatccgcc gcgccctccc cagggtcttc tttcgtttgt gccttccctg 60

tccaggtcac agcagcacca ccagccccgc ttgtacgtcg taggttcc 108

<210> 197

<211> 108

<212> DNA

<213>artificial sequence

<400> 197

gactacatgg gacatacgtg gacagctgcc ctttcaaaag cccagggaac tcagaaggac 60

ctctctcccc agagtggaga atgggttcct ggttacgtcg taggttcc 108

<210> 198

<211> 108

<212> DNA

<213>artificial sequence

<400> 198

gactacatgg gacataggaa tacgctggcg ttcaggaaac ggatgacttg taagctctgt 60

atttgagttg tgtagactct gcagcatcac atatacgtcg taggttcc 108

<210> 199

<211> 108

<212> DNA

<213>artificial sequence

<400> 199

gactacatgg gacattacgt gatgtggact agcaaaaaaa agaaagatac catattaaaa 60

taagaggagg atacaaatga agggatgaaa gtatacgtcg taggttcc 108

<210> 200

<211> 108

<212> DNA

<213>artificial sequence

<400> 200

gactacatgg gacatctcat cccctctccc actctttaaa aaaatactat tatctgatta 60

tgaacaaaga accttctctt taatgcatcc tactacgtcg taggttcc 108

<210> 201

<211> 108

<212> DNA

<213>artificial sequence

<400> 201

gactacatgg gacatgtgaa tcaaagaaag atgacctgat gcatgtcatc agttaattta 60

ccgtgtttga gatctcttac ttattcaatc aaatacgtcg taggttcc 108

<210> 202

<211> 108

<212> DNA

<213>artificial sequence

<400> 202

gactacatgg gacatggtgg tctgtgtcca aaatggctac aaagcaacct aggaaagtca 60

caaagtaaga gaaaagcctg gtccttttct ttttacgtcg taggttcc 108

<210> 203

<211> 108

<212> DNA

<213>artificial sequence

<400> 203

gactacatgg gacatctatt acaccatgaa gagcagtcat tcaaatggaa agggttgaaa 60

gctttgcaaa gaaaatattc tccttgagta acatacgtcg taggttcc 108

<210> 204

<211> 108

<212> DNA

<213>artificial sequence

<400> 204

gactacatgg gacatatcct gaaatatcta taaagaatgt tctagagaat tcaagatgca 60

caatttgaaa gaaaatgtca acactgtcta aaatacgtcg taggttcc 108

<210> 205

<211> 108

<212> DNA

<213>artificial sequence

<400> 205

gactacatgg gacattgtta ctatcagttt cagtgacaca acatcagagg caatctgata 60

ttgacaatgg taaaaatatg cagacgccaa tattacgtcg taggttcc 108

<210> 206

<211> 108

<212> DNA

<213>artificial sequence

<400> 206

gactacatgg gacataatat taggagggtg cagttagttg acagtagagc ctataagtat 60

tcatctaagt tttctctgtt tgaagagaag aaatacgtcg taggttcc 108

<210> 207

<211> 108

<212> DNA

<213>artificial sequence

<400> 207

gactacatgg gacatgcctt tgtctgtcca gcaggcacat gaagattgtg gtggtgggca 60

ggagaggcta aggggaaaca gaggtgaggg gagtacgtcg taggttcc 108

<210> 208

<211> 108

<212> DNA

<213>artificial sequence

<400> 208

gactacatgg gacatcacag gcaaatcaaa actctactta atgacttgga gaccagcaga 60

tgaggattag gggaggcaac atttatttgg agatacgtcg taggttcc 108

<210> 209

<211> 108

<212> DNA

<213>artificial sequence

<400> 209

gactacatgg gacatactac aaaaatgctg cttattccac tccaaatcat tagctcatca 60

tccatgtaag gagcttcaag aagaatatcg gtatacgtcg taggttcc 108

<210> 210

<211> 108

<212> DNA

<213>artificial sequence

<400> 210

gactacatgg gacataccat atttgccttt ttatttgatt gactacatcc aacttgttta 60

gcaatctacc cagctaaccg cgttcactca cgttacgtcg taggttcc 108

<210> 211

<211> 108

<212> DNA

<213>artificial sequence

<400> 211

gactacatgg gacatcacgt ttaactatat agcattgctc tgtatttgtc tttcctttca 60

gactgagcag tactggtgaa atactatttg tcctacgtcg taggttcc 108

<210> 212

<211> 108

<212> DNA

<213>artificial sequence

<400> 212

gactacatgg gacatctttc atcagtcctt acgggaactc ctctaagccc ctaatccctt 60

ttcctgactt tccttgtgcc cttcacaccc tgctacgtcg taggttcc 108

<210> 213

<211> 108

<212> DNA

<213>artificial sequence

<400> 213

gactacatgg gacatgttga tgttttaggg gaaattaaaa gaacattcgg ttttctgagt 60

ctccttccgg ggaggcgtgg tggtaactgg ttttacgtcg taggttcc 108

<210> 214

<211> 108

<212> DNA

<213>artificial sequence

<400> 214

gactacatgg gacatccatg atacacactt gaatctcaag ccccaagtgt ggcttcctcc 60

accccgtgat taagtacggc tgcctgttgt cggtacgtcg taggttcc 108

<210> 215

<211> 108

<212> DNA

<213>artificial sequence

<400> 215

gactacatgg gacatttttt cctggggctt tagacaatct gtggaaaagg aacacaatca 60

gcgccagcga tgtgctccgt ggcaccattg atctacgtcg taggttcc 108

<210> 216

<211> 108

<212> DNA

<213>artificial sequence

<400> 216

gactacatgg gacatgacgc agagtgtctg cgcatgggtc ctgttgagtg cgtgatctct 60

tgtgtgcgga tttaattctg tcgagtgggt atctacgtcg taggttcc 108

<210> 217

<211> 108

<212> DNA

<213>artificial sequence

<400> 217

gactacatgg gacatgtatc tcttgtgtgc aggtttcgtt ggtgagcgtg aacgttcgtg 60

cacacccctt gctgtgtgtg cgtgtcacga ctctacgtcg taggttcc 108

<210> 218

<211> 108

<212> DNA

<213>artificial sequence

<400> 218

gactacatgg gacatgagac atagtcttaa aggtagcatt cagccggtga gatatctttc 60

tttctggccg aggctttcct ggctttataa atttacgtcg taggttcc 108

<210> 219

<211> 108

<212> DNA

<213>artificial sequence

<400> 219

gactacatgg gacatacgaa aatatttaat tcatggtatt caggtttcct gtgcacggag 60

cctctgcgct gcgttactgt atttattcat cggtacgtcg taggttcc 108

<210> 220

<211> 108

<212> DNA

<213>artificial sequence

<400> 220

gactacatgg gacatccctc ccccactggg gacctctcct aggggagggg tctggtgtcc 60

tcagctggct cctctccacc ttcgcttcta ccctacgtcg taggttcc 108

<210> 221

<211> 108

<212> DNA

<213>artificial sequence

<400> 221

gactacatgg gacatccttg cttcctccaa cacccccacc ggctttctcc tgtggcttaa 60

aattgaccta gtcagtcctg ccagtgggtc tactacgtcg taggttcc 108

<210> 222

<211> 108

<212> DNA

<213>artificial sequence

<400> 222

gactacatgg gacattggaa gcagcagcta tcccgaagaa gaaacggtgt ctagaaatga 60

atcacattac tgctgaggtc agcgcttccg aagtacgtcg taggttcc 108

<210> 223

<211> 108

<212> DNA

<213>artificial sequence

<400> 223

gactacatgg gacatcgaag acaggctaca gaggtgggag acgggagtct gagtggccca 60

ctggggtcgc gtgccctcag agcccggaga aggtacgtcg taggttcc 108

<210> 224

<211> 108

<212> DNA

<213>artificial sequence

<400> 224

gactacatgg gacataaaat actgagtaat ttgggtacaa tcagaaatct ctgccggctg 60

atgacgtgat ttccaaatga agaccacccg ggctacgtcg taggttcc 108

<210> 225

<211> 108

<212> DNA

<213>artificial sequence

<400> 225

gactacatgg gacatttgtc tgttttgaac actatactgt gaagtccacg aggtagaggc 60

acagtgacag caagcctcac tctacatctt cagtacgtcg taggttcc 108

<210> 226

<211> 108

<212> DNA

<213>artificial sequence

<400> 226

gactacatgg gacatcagag tcctctgtca tcattttgcc tctctcctaa gtgacaggac 60

tgagtgcaga cttggtgttt gtgggtgagg cattacgtcg taggttcc 108

<210> 227

<211> 108

<212> DNA

<213>artificial sequence

<400> 227

gactacatgg gacatactta cccctctgtc tatattctat tttccagcca aatatgaggt 60

gacgtttaag ctgctacttg aaagagaagt gggtacgtcg taggttcc 108

<210> 228

<211> 108

<212> DNA

<213>artificial sequence

<400> 228

gactacatgg gacattggag cagtggcact tggacagcaa tccaggatca ttgatgctag 60

ccacagcact cctggggtgt gtgtatagag gggtacgtcg taggttcc 108

<210> 229

<211> 108

<212> DNA

<213>artificial sequence

<400> 229

gactacatgg gacatggtgg tgcagtggga gtccactcac ttttcaggag gctttggaaa 60

gctgggggtt gttccctggc tttttctccc acttacgtcg taggttcc 108

<210> 230

<211> 108

<212> DNA

<213>artificial sequence

<400> 230

gactacatgg gacatttatc ctgcagccca tcaggatctg ggcactgaac tgcaactctt 60

agtgcagtca gtacatccag tgaagcagaa gcatacgtcg taggttcc 108

<210> 231

<211> 108

<212> DNA

<213>artificial sequence

<400> 231

gactacatgg gacatgttct gaagtctaca ggttttcagt ctgtgttctg gaagattttt 60

caaggtcaga ggcagtgaat gggagggcag ggatacgtcg taggttcc 108

<210> 232

<211> 108

<212> DNA

<213>artificial sequence

<400> 232

gactacatgg gacattgtat gaaagggacg gttgtggtga agtttgtcag gcacagtaag 60

catgctgagt ggcggggatc agaactctcc tattacgtcg taggttcc 108

<210> 233

<211> 108

<212> DNA

<213>artificial sequence

<400> 233

gactacatgg gacatcctgt caatcacgca ccgtctgtct ctggcagtgg caatgttgtt 60

gctgagtcag attgctggtg gcagggttgt cattacgtcg taggttcc 108

<210> 234

<211> 108

<212> DNA

<213>artificial sequence

<400> 234

gactacatgg gacattaggc agtttctaag actttgggaa aatccctggg agttcctcat 60

aacacaaaat gccttgggct gtacaagcct aggtacgtcg taggttcc 108

<210> 235

<211> 108

<212> DNA

<213>artificial sequence

<400> 235

gactacatgg gacatgatac acggacctga acaaaggtag ggggcatggg gatggagatg 60

aaaggaaacc atcagaaaag aaggggccaa agatacgtcg taggttcc 108

<210> 236

<211> 108

<212> DNA

<213>artificial sequence

<400> 236

gactacatgg gacataaaga gggaggttat cttggcagaa gaaagacatt attcatcttg 60

gttggttgga aatgtgggtc tggagctcat aagtacgtcg taggttcc 108

<210> 237

<211> 108

<212> DNA

<213>artificial sequence

<400> 237

gactacatgg gacatgaatg cttaaacgat ggccagattt tactctgagt gatactaccc 60

cctggaagtg gggggaaaaa aagaaggagg gaatacgtcg taggttcc 108

<210> 238

<211> 108

<212> DNA

<213>artificial sequence

<400> 238

gactacatgg gacatagcct gggccctgga gggcgcagcc ggtgcagtct gcaggaggct 60

ctgtgtttca ataactgtgg tgtccccaca ccctacgtcg taggttcc 108

<210> 239

<211> 108

<212> DNA

<213>artificial sequence

<400> 239

gactacatgg gacatggggc aagagagaat ttaaggggtg gcaaggggcc actgtcccgc 60

atcttgggga cgttgtaggc aggcaggtag agatacgtcg taggttcc 108

<210> 240

<211> 108

<212> DNA

<213>artificial sequence

<400> 240

gactacatgg gacataattt ctctgtgctc tgttttgagg ggatggtcct ggtactggag 60

tagcagagcc aggtccctgc ttcttgcata acctacgtcg taggttcc 108

<210> 241

<211> 108

<212> DNA

<213>artificial sequence

<400> 241

gactacatgg gacataggga aactgactct agattatgga aataaacatt gtcttcaagg 60

gatagccagc aacatcaggc tcagggctag tgatacgtcg taggttcc 108

<210> 242

<211> 108

<212> DNA

<213>artificial sequence

<400> 242

gactacatgg gacatgggat tagtgtcagc cactcctctc cctccccctg gaacatcctt 60

gtgggtaggg gacgatgaca cttggtgact ggttacgtcg taggttcc 108

<210> 243

<211> 108

<212> DNA

<213>artificial sequence

<400> 243

gactacatgg gacatttgct gaatccagac tggccccagg ccagactctc aaacaagggc 60

caggacatgg ctcactctgc ctgttgtcca tcgtacgtcg taggttcc 108

<210> 244

<211> 108

<212> DNA

<213>artificial sequence

<400> 244

gactacatgg gacatcacga tccagcccaa ggtaatgaag gtggggaagc ctcggaatgc 60

aagagacagg gtggagatga gaagacatgt cagtacgtcg taggttcc 108

<210> 245

<211> 108

<212> DNA

<213>artificial sequence

<400> 245

gactacatgg gacatatcca ctggagtgac cgaaggccag gtgtgctccc agaggcaggg 60

ataacagtgg tgagcaagga gccaccatgc tcatacgtcg taggttcc 108

<210> 246

<211> 108

<212> DNA

<213>artificial sequence

<400> 246

gactacatgg gacatgaata gcgcctctcc atccagttct catctttgag aacgattctt 60

gggatgaaag ccacgacgtt ggatatgaac aactacgtcg taggttcc 108

<210> 247

<211> 108

<212> DNA

<213>artificial sequence

<400> 247

gactacatgg gacattctgg acatctctgg gagtgaaaag ggagttctat gctcagccac 60

ctgagtttgt ggagctcgtt tcaatctcag tcttacgtcg taggttcc 108

<210> 248

<211> 108

<212> DNA

<213>artificial sequence

<400> 248

gactacatgg gacatataaa acacacaaaa agcctctgtg gaggacagat gtgtagtgtc 60

tgttgttcta caccattcat tccagagttt agctacgtcg taggttcc 108

<210> 249

<211> 108

<212> DNA

<213>artificial sequence

<400> 249

gactacatgg gacattacat tctgtgaatg aatatggaca atcaccatgt ggcaaaggtt 60

atcccttctc cctctgaccc ctggcaattg tgatacgtcg taggttcc 108

<210> 250

<211> 108

<212> DNA

<213>artificial sequence

<400> 250

gactacatgg gacatccaat ggctcttcct ttgctgccca gggcttttcc tgtctcctgt 60

gtgtgagttg gggtgggtgt gtggggtact ggctacgtcg taggttcc 108

<210> 251

<211> 108

<212> DNA

<213>artificial sequence

<400> 251

gactacatgg gacatctttg ccagctcctt aacaaagggt caaggtggag catgaagaat 60

aatgggaaaa ggggtgaggg atgacatcgt gtgtacgtcg taggttcc 108

<210> 252

<211> 108

<212> DNA

<213>artificial sequence

<400> 252

gactacatgg gacatactgg cattcaggca gctttctatc tcccagccct caagggaagg 60

aatgatcatg tctaggataa aggccagatt ttttacgtcg taggttcc 108

<210> 253

<211> 108

<212> DNA

<213>artificial sequence

<400> 253

gactacatgg gacatctctg tctcagtgtt ggtgttgggg ccacgagggg gtgatgggga 60

cattgatgag ctggagtgaa gagcaccaaa gcctacgtcg taggttcc 108

<210> 254

<211> 108

<212> DNA

<213>artificial sequence

<400> 254

gactacatgg gacatatttg tttgcgttgt ctaaaattat cctctgattt cattcggtgc 60

ctgcgtcagg agggagaaac atgggaaggt tgttacgtcg taggttcc 108

<210> 255

<211> 108

<212> DNA

<213>artificial sequence

<400> 255

gactacatgg gacatgtccc agctggcgtg ctggacgaat tcggaaacac cttggtctta 60

cttccctaaa atctccatct acctgagaga cgatacgtcg taggttcc 108

<210> 256

<211> 108

<212> DNA

<213>artificial sequence

<400> 256

gactacatgg gacattgatt cttccctact gcctggtgtc tgagcctctg tcttgacagg 60

ctgcctcggc tcctgcccat cctgctttca tcctacgtcg taggttcc 108

<210> 257

<211> 108

<212> DNA

<213>artificial sequence

<400> 257

gactacatgg gacattgcat tctggctgcc ggtgcagagg gctgtcatgt aagcccacca 60

caataataga accagccagt ctgctcctcc agctacgtcg taggttcc 108

<210> 258

<211> 108

<212> DNA

<213>artificial sequence

<400> 258

gactacatgg gacatggact cagaggagct gcggctgagc tcctcggggc cctcgacagc 60

ctgggaaact ccagagtcca gctgtgagag cagtacgtcg taggttcc 108

<210> 259

<211> 108

<212> DNA

<213>artificial sequence

<400> 259

gactacatgg gacatcgcac agcggccgag gggagacgtg accggcgccc tcacctgtcc 60

tgcagttgtg gaactccagc gcgctctcga tgctacgtcg taggttcc 108

<210> 260

<211> 108

<212> DNA

<213>artificial sequence

<400> 260

gactacatgg gacattgtga ggacacggcc cagccccacc gtcgtcttcc agccccacca 60

tcgtcttcca gccccaccgt cgtcttccag tcctacgtcg taggttcc 108

<210> 261

<211> 108

<212> DNA

<213>artificial sequence

<400> 261

gactacatgg gacatctcac tgaccctttg ctcatcttct ctgtctgtgc tttcgtagat 60

tgtcctccag tcagtgacta cgtggtcctc aactacgtcg taggttcc 108

<210> 262

<211> 108

<212> DNA

<213>artificial sequence

<400> 262

gactacatgg gacattctca tttcatttcc aaagtacacc aggactgagg ggagggacac 60

tcttttccct ttttggaaac tgaggcctga gagtacgtcg taggttcc 108

<210> 263

<211> 108

<212> DNA

<213>artificial sequence

<400> 263

gactacatgg gacatggggt acttaatcca gatactgggc caatccatct ttaagatatg 60

tgcctttatt caaatggtgt acaccactta ctctacgtcg taggttcc 108

<210> 264

<211> 108

<212> DNA

<213>artificial sequence

<400> 264

gactacatgg gacatatcgg tcacacctgg aaccctgctt aatggtcgtg aagacgttga 60

tggatgtctg ctgttgcctc ttcttggcta ggttacgtcg taggttcc 108

<210> 265

<211> 108

<212> DNA

<213>artificial sequence

<400> 265

gactacatgg gacatgcttt tcctggggcc agagtaggtg gagctgagct ccccaaagct 60

tccagtttgc tgcctgctcc cacccgcctc gcctacgtcg taggttcc 108

<210> 266

<211> 108

<212> DNA

<213>artificial sequence

<400> 266

gactacatgg gacattggtg ttatgctcag ctggaccccc acccccggtg tggaaagctc 60

atgttatact ccactggccc ccacccccgg cgttacgtcg taggttcc 108

<210> 267

<211> 108

<212> DNA

<213>artificial sequence

<400> 267

gactacatgg gacatccccc gcctgtggtg agttcctcga atggtgcatg gggactagga 60

ctaataggac ccatagcctg tgtgagcctc tggtacgtcg taggttcc 108

<210> 268

<211> 108

<212> DNA

<213>artificial sequence

<400> 268

gactacatgg gacatgacta ggactaatag gacccatagc ctgtgtgagc ctctggggca 60

aagctcctgc agccgtggag tcgcaggctg ctatacgtcg taggttcc 108

<210> 269

<211> 108

<212> DNA

<213>artificial sequence

<400> 269

gactacatgg gacatgagag atgggtagat tttcctactt gggtccttct gtggacaggt 60

actaggtgct gctttacgcc cagtgacttg tgatacgtcg taggttcc 108

<210> 270

<211> 108

<212> DNA

<213>artificial sequence

<400> 270

gactacatgg gacatggacg gggggttgga caattcctta tagggaagga ggtggtgaat 60

acgcgggaac cataattcta ccagagaagt tgatacgtcg taggttcc 108

<210> 271

<211> 108

<212> DNA

<213>artificial sequence

<400> 271

gactacatgg gacataccat cccaaggaga gacttcatct gagatcaagc cagcaaatgt 60

ttccccggcg cccacatgag aaaggcgccg tgttacgtcg taggttcc 108

<210> 272

<211> 108

<212> DNA

<213>artificial sequence

<400> 272

gactacatgg gacatgtcag actctgctct ctgcctggaa tttccttcct ggtttcaccc 60

attccttgcc acctccctcc cagctaaccc ggatacgtcg taggttcc 108

<210> 273

<211> 108

<212> DNA

<213>artificial sequence

<400> 273

gactacatgg gacataacag aggttcatta gcaacagcaa agggcttaga gccaacagtg 60

cctgccaccc tccaccctta gagaattaaa ccatacgtcg taggttcc 108

<210> 274

<211> 108

<212> DNA

<213>artificial sequence

<400> 274

gactacatgg gacatccaac agtgcctgcc accctccacc cttagagaat taaaccagtg 60

ctggagtaga gcgtcaccag gtggtggaag gactacgtcg taggttcc 108

<210> 275

<211> 108

<212> DNA

<213>artificial sequence

<400> 275

gactacatgg gacatgtcgc agagccttgg gtggactgtt tattagagga atattttatg 60

caggtaagag tcttgcagag caatcaagcc tcctacgtcg taggttcc 108

<210> 276

<211> 108

<212> DNA

<213>artificial sequence

<400> 276

gactacatgg gacataggta gctggtgact gcctggccag caggggaaac cgaggcaccc 60

aggatggctt cacagccaga cctggggaga gtgtacgtcg taggttcc 108

<210> 277

<211> 108

<212> DNA

<213>artificial sequence

<400> 277

gactacatgg gacataccca gaaaggccgt tccagctcct tctggactcc tcctcctcct 60

ccccagtgtt ctctgtctct ctgcccaccc cgctacgtcg taggttcc 108

<210> 278

<211> 108

<212> DNA

<213>artificial sequence

<400> 278

gactacatgg gacatgaggc cacctccatc ctctccttct gcctccttgg gaagcccctg 60

tagttccctc tgtaagatgc tctgtccctt tgttacgtcg taggttcc 108

<210> 279

<211> 108

<212> DNA

<213>artificial sequence

<400> 279

gactacatgg gacatacctg gagctgctga ggcttgagcc ctggaggccc tggccggttc 60

cctgagttgg tacagggcct gtgtgtgcac gtgtacgtcg taggttcc 108

<210> 280

<211> 108

<212> DNA

<213>artificial sequence

<400> 280

gactacatgg gacatagctc cctctcctgc tgcaatgcag cctccacgtg caatgcagcc 60

tccacgtgca atgcagcctc cacgtgcaat gcatacgtcg taggttcc 108

<210> 281

<211> 108

<212> DNA

<213>artificial sequence

<400> 281

gactacatgg gacattgggc taggaatgtg aacaggaaac tttcagtgtg gacggccatg 60

catttaggcc atagtgggtt cccagcagcc gtctacgtcg taggttcc 108

<210> 282

<211> 108

<212> DNA

<213>artificial sequence

<400> 282

gactacatgg gacattgaag ggaggttgaa ggtctcccat tatatgtgac tcctcttcca 60

gcacccaagt gagccctggg gtggggtgtg tcttacgtcg taggttcc 108

<210> 283

<211> 108

<212> DNA

<213>artificial sequence

<400> 283

gactacatgg gacatacatc acggctgctg aagttttgtt gtccgatgca cccagctcta 60

tagggagtgt gggccatcct tgaaaacgga gtatacgtcg taggttcc 108

<210> 284

<211> 108

<212> DNA

<213>artificial sequence

<400> 284

gactacatgg gacattgggt ggggccctgg atcttgatgt tacctggccc ttgcctgtgt 60

gccaggtgga ctggggactg tggatgtctg ggctacgtcg taggttcc 108

<210> 285

<211> 108

<212> DNA

<213>artificial sequence

<400> 285

gactacatgg gacatcagcg tcaacagttg catttgtttt tttatccctt cacaggttcc 60

ctcccactta ctacgctcca agattaccaa gcctacgtcg taggttcc 108

<210> 286

<211> 108

<212> DNA

<213>artificial sequence

<400> 286

gactacatgg gacatagcaa gacagcgagc tggcccctgc cgcacctgtg ggctgctggg 60

tggagagtcc tggtttcaca gcatgtccaa cgttacgtcg taggttcc 108

<210> 287

<211> 108

<212> DNA

<213>artificial sequence

<400> 287

gactacatgg gacataatgt tttccttgaa aaaacctctt gatgatgaag ccggtttaac 60

tggtgttgac caatggggtg tcagctttga aggtacgtcg taggttcc 108

<210> 288

<211> 108

<212> DNA

<213>artificial sequence

<400> 288

gactacatgg gacatatctt tcccccagat gtcatcagtg cctgcttccc tccctgtgga 60

gggctctgag ggccgacccc caaagtccat ctttacgtcg taggttcc 108

<210> 289

<211> 108

<212> DNA

<213>artificial sequence

<400> 289

gactacatgg gacatcctca acctccttgt ttctaggttg cagctccatt acgtgctctg 60

caagtgaaag atgctgaggt ggacactaac gtctacgtcg taggttcc 108

<210> 290

<211> 108

<212> DNA

<213>artificial sequence

<400> 290

gactacatgg gacataagca tgctgggtga taagaaggca ttctgggggg ccagggtctc 60

ttgtaacaaa ttcaggaagc atctcccaga ctttacgtcg taggttcc 108

<210> 291

<211> 108

<212> DNA

<213>artificial sequence

<400> 291

gactacatgg gacatcacgg ggctgggggt gctacctcct acaggacaag gtcagaggac 60

actgccccag actactacag gcaggcacca aactacgtcg taggttcc 108

<210> 292

<211> 108

<212> DNA

<213>artificial sequence

<400> 292

gactacatgg gacatcagca ttcgccccag gagtgaactg gaagtgccag cccgggccag 60

ctgggtcaag gcccactgcc tgaggctgcc agctacgtcg taggttcc 108

<210> 293

<211> 108

<212> DNA

<213>artificial sequence

<400> 293

gactacatgg gacataggca agagggcagc atcttgatag tgagggttgg tcacccggga 60

accacagtaa cgtctgcagg gaccttcccc cagtacgtcg taggttcc 108

<210> 294

<211> 108

<212> DNA

<213>artificial sequence

<400> 294

gactacatgg gacatcatag ggggatgtgt gaggaccata gaggggacat gtgaggatca 60

tgggggacat gtgaggatca tgggggggac gtgtacgtcg taggttcc 108

<210> 295

<211> 108

<212> DNA

<213>artificial sequence

<400> 295

gactacatgg gacattctcc cttgaagaaa ctgacttctg tgactggaaa gatcgggtgc 60

tcctgcctag aaagatccaa gccgtccaac gtatacgtcg taggttcc 108

<210> 296

<211> 108

<212> DNA

<213>artificial sequence

<400> 296

gactacatgg gacatatcgg gtgctcctgc ctagaaagat ccaagccgtc caacgtacag 60

ctgggctttc atgttaaacc cagggagtgg ggttacgtcg taggttcc 108

<210> 297

<211> 108

<212> DNA

<213>artificial sequence

<400> 297

gactacatgg gacattctca gttgcgtggg gaccagaggg tgctggagaa acaaaccaga 60

cgcagctgaa ggcagtcagg gcagggcgca atctacgtcg taggttcc 108

<210> 298

<211> 108

<212> DNA

<213>artificial sequence

<400> 298

gactacatgg gacatggagt gtgcacagag catgaggcag gagtgcagag catgaggcgg 60

gggatgcaca gagcatgagg caggagtgca gagtacgtcg taggttcc 108

<210> 299

<211> 108

<212> DNA

<213>artificial sequence

<400> 299

gactacatgg gacatgggtg aaggctggct agcccgtagg aggtcaggct cacagaggca 60

gctcatggag atccttggag gggcgcacct tactacgtcg taggttcc 108

<210> 300

<211> 108

<212> DNA

<213>artificial sequence

<400> 300

gactacatgg gacatcactt tctttcacat agcctgcccc tttcccatgt tgcctccctt 60

cctgatgctg ccccaccagc ccccattcct tcctacgtcg taggttcc 108

<210> 301

<211> 108

<212> DNA

<213>artificial sequence

<400> 301

gactacatgg gacatgagac ctgtacatgg tcacacccct ttgcttctga agccctgctt 60

caaatctcag ctccccagag agctctgcca cagtacgtcg taggttcc 108

<210> 302

<211> 108

<212> DNA

<213>artificial sequence

<400> 302

gactacatgg gacatgcatt tgccactgct gaaaccggag cccccagaag cttttttggg 60

cacttgtcag gagcctgttg ctataggaac tgttacgtcg taggttcc 108

<210> 303

<211> 108

<212> DNA

<213>artificial sequence

<400> 303

gactacatgg gacatagtta ccatgggtgc atgtaatata aaccccgcaa agactgttta 60

caattccgtg tgggtgtagg caaaaaaaaa aggtacgtcg taggttcc 108

<210> 304

<211> 108

<212> DNA

<213>artificial sequence

<400> 304

gactacatgg gacatacata aaaagcagga tgagacagac agaagccatg gccgtgagcc 60

tgaatctcac taacctctct cattggcttt ccatacgtcg taggttcc 108

<210> 305

<211> 108

<212> DNA

<213>artificial sequence

<400> 305

gactacatgg gacattgaac tagggaagga gttgttgagt tgctccatca cctcctctaa 60

ccctgtgctt gtgtcctggg gaggactgag aagtacgtcg taggttcc 108

<210> 306

<211> 108

<212> DNA

<213>artificial sequence

<400> 306

gactacatgg gacattccag cagctgcctt agcctgtgta actgtgactc cagctgttta 60

ttgtggtctt ccaggatttg catcctggct tcctacgtcg taggttcc 108

<210> 307

<211> 108

<212> DNA

<213>artificial sequence

<400> 307

gactacatgg gacatgaagt aagtacacct tcagtcagat tctgcacacg ggtcttctgg 60

gtgtgcaccg atgtatgtaa ctgtccacat taatacgtcg taggttcc 108

<210> 308

<211> 108

<212> DNA

<213>artificial sequence

<400> 308

gactacatgg gacattcatt gtcaggaaca ttttgtaaaa agagatggga tacttacatg 60

ctctcattag gagagatgct atcatttaga taatacgtcg taggttcc 108

<210> 309

<211> 108

<212> DNA

<213>artificial sequence

<400> 309

gactacatgg gacatttggt ggtagcagca cccttcagca aaaaaagtac tcacgcagaa 60

tctactggcc agaagttgat cagagtaacg ggatacgtcg taggttcc 108

<210> 310

<211> 108

<212> DNA

<213>artificial sequence

<400> 310

gactacatgg gacatttacc ggagtgcaat attccaccat gggatagtgc attttatggc 60

cttttgcaac tcgaccagaa aaaaagcagc ttttacgtcg taggttcc 108

<210> 311

<211> 108

<212> DNA

<213>artificial sequence

<400> 311

gactacatgg gacattggac tggggttcca gtctcatcca gtctaggaag agggccgctt 60

cgatctctgg cttattatta gcctgcaaag acatacgtcg taggttcc 108

<210> 312

<211> 108

<212> DNA

<213>artificial sequence

<400> 312

gactacatgg gacatgcaac ttcacccaac tgtcttggaa tttggataga atcatgcaga 60

aggaggccca gcctgcgctg gtcacaaaat ccttacgtcg taggttcc 108

<210> 313

<211> 108

<212> DNA

<213>artificial sequence

<400> 313

gactacatgg gacattaaaa acatgccata cgtacgtatc ataaacattc agcagccagt 60

tcagacacat atccacgcag agagggacgt tgatacgtcg taggttcc 108

<210> 314

<211> 108

<212> DNA

<213>artificial sequence

<400> 314

gactacatgg gacatttgtt ccaagagaga aaatgcatcg tggtgtggct cttgcagaca 60

gagggcaatt atagcttgca cctgtcctct ctgtacgtcg taggttcc 108

<210> 315

<211> 108

<212> DNA

<213>artificial sequence

<400> 315

gactacatgg gacatcttgg aggaaacatg gccatgtcct tacctaaaga ctggtagagc 60

tctgtcattt tgggatggtc ccagcaagtt gtttacgtcg taggttcc 108

<210> 316

<211> 108

<212> DNA

<213>artificial sequence

<400> 316

gactacatgg gacatggaga aagtctcctg gtggcaagcc ttgaacagcc agcaagtggc 60

aggaattcca cacaaagaca gggatccata aaatacgtcg taggttcc 108

<210> 317

<211> 108

<212> DNA

<213>artificial sequence

<400> 317

gactacatgg gacatggcgt tgcctggctg agctaatggg aagacctacg tagggtatct 60

agagttgact ttttatgttt acattaaaag gggtacgtcg taggttcc 108

<210> 318

<211> 108

<212> DNA

<213>artificial sequence

<400> 318

gactacatgg gacatccaaa agcactttag aaggatggaa tcttatcatt tactccctga 60

atcactatca tcagtacagt tggtgtagaa cggtacgtcg taggttcc 108

<210> 319

<211> 108

<212> DNA

<213>artificial sequence

<400> 319

gactacatgg gacatccatc tggtgttcag gtcttccaga gtgctgaggt tatacggtga 60

gagctgaatg cccaaagtgg taagctggcg agctacgtcg taggttcc 108

<210> 320

<211> 108

<212> DNA

<213>artificial sequence

<400> 320

gactacatgg gacatgagtc aatgaggaga tcgcccacgg gctgccagga tcccttgatc 60

acctcagctt ggcgcagctt gaggtccagc tcatacgtcg taggttcc 108

<210> 321

<211> 108

<212> DNA

<213>artificial sequence

<400> 321

gactacatgg gacatgacct cctcagcctg ctttcgtaga agccgagtga cattctgggc 60

tctctcctca ggaggcagct ctaaattggc aattacgtcg taggttcc 108

<210> 322

<211> 108

<212> DNA

<213>artificial sequence

<400> 322

gactacatgg gacatgctgg aaagtcgcct ccaataggtg cctgccggct taattcatca 60

tctttcagct gtagccacac cagaagttcc tgctacgtcg taggttcc 108

<210> 323

<211> 108

<212> DNA

<213>artificial sequence

<400> 323

gactacatgg gacatccaaa cgtctttgta acaggactgc atcatcggaa ccttccaggg 60

atctcaggat tttttggctg ttttcatcca ggttacgtcg taggttcc 108

<210> 324

<211> 108

<212> DNA

<213>artificial sequence

<400> 324

gactacatgg gacataatct aatctttaaa aatgcctggc ctagggtgga ttacccccgg 60

aggaaactgt caaaccgcct taagcaatcc cgatacgtcg taggttcc 108

<210> 325

<211> 108

<212> DNA

<213>artificial sequence

<400> 325

gactacatgg gacatccctt ggagtcttct aggagccttt ccttacgggt agcatcctgt 60

aggacattgg cagttgtttc agcttctgta agctacgtcg taggttcc 108

<210> 326

<211> 108

<212> DNA

<213>artificial sequence

<400> 326

gactacatgg gacatatgta attcatacct tttatgaatg cttctccaag aggcattgat 60

attctctgtt atcatgtgga cttttctggt atctacgtcg taggttcc 108

<210> 327

<211> 108

<212> DNA

<213>artificial sequence

<400> 327

gactacatgg gacattgtat agggaccctc cttccatgac tcaagcttgg ctctggcctg 60

tcctaagacc tgctcagctt cttccttagc ttctacgtcg taggttcc 108

<210> 328

<211> 108

<212> DNA

<213>artificial sequence

<400> 328

gactacatgg gacatggtct tttaaatctc tccttttggg gtggtggtgg ggggcgaaat 60

aactaaacag ctaccaaaaa gctcattaag tattacgtcg taggttcc 108

<210> 329

<211> 108

<212> DNA

<213>artificial sequence

<400> 329

gactacatgg gacatatcgt cttatgagtt atgcatgtgc aagaaatgtt ctgcatgttt 60

gtgtgcgtga gagagacaca gagagagagc gagtacgtcg taggttcc 108

<210> 330

<211> 108

<212> DNA

<213>artificial sequence

<400> 330

gactacatgg gacataggta cctccaacat caaggaagat ggcatttcta gtttggagat 60

ggcagtttcc ttagtaacca caggttgtgt cactacgtcg taggttcc 108

<210> 331

<211> 108

<212> DNA

<213>artificial sequence

<400> 331

gactacatgg gacattccaa tagtggtcag tccaggagct aggtcaggct gctttgccct 60

cagctcttga agtaaacggt ttaccgcctt ccatacgtcg taggttcc 108

<210> 332

<211> 108

<212> DNA

<213>artificial sequence

<400> 332

gactacatgg gacatagggg atggacggca gagagaatgt attccaagag ggaacatgga 60

acatgcaaag acaggaagca gaagaagtac ggatacgtcg taggttcc 108

<210> 333

<211> 108

<212> DNA

<213>artificial sequence

<400> 333

gactacatgg gacatgtgtg actgatttcc aatagcctcc aacttcttta gcctgccttt 60

atctaaaaaa tggttcttaa caatctgacg ctatacgtcg taggttcc 108

<210> 334

<211> 108

<212> DNA

<213>artificial sequence

<400> 334

gactacatgg gacatttacc ttgacttgct caagcttttc ttttagttgc tgctcttttc 60

caggttcaag tgggatacta gcaatgttat ctgtacgtcg taggttcc 108

<210> 335

<211> 108

<212> DNA

<213>artificial sequence

<400> 335

gactacatgg gacatacctc ctgccaccgc agattcaggc ttcccaattt ttcctgtaga 60

atactggcat ctgtttttga ggattgctga atttacgtcg taggttcc 108

<210> 336

<211> 108

<212> DNA

<213>artificial sequence

<400> 336

gactacatgg gacatggacg tggtggggag aaatcttgta aagtcaacaa tccattttct 60

agttactgtc tagtcacgca ttataaatga cactacgtcg taggttcc 108

<210> 337

<211> 108

<212> DNA

<213>artificial sequence

<400> 337

gactacatgg gacattgaaa gtattcaccc acaccgtaat ccgctggttt attccacctt 60

tatgaaactc ctattacatt tgtccaccct gtgtacgtcg taggttcc 108

<210> 338

<211> 108

<212> DNA

<213>artificial sequence

<400> 338

gactacatgg gacattcaaa gacttacctt aagataccat ttgtatttag catgttccca 60

attctcagga atttgtgtct ttctgagaaa ctgtacgtcg taggttcc 108

<210> 339

<211> 108

<212> DNA

<213>artificial sequence

<400> 339

gactacatgg gacattattc ttgtccttgg ttcaaggatc gaacactcaa ggagacctaa 60

atattttgga ataggagcca ggataacctg tcctacgtcg taggttcc 108

<210> 340

<211> 108

<212> DNA

<213>artificial sequence

<400> 340

gactacatgg gacatacagg cgttgcactt tgcaatgctg ctgtcttctt gctatgaata 60

atgtcaatcc gacctgagct ttgttgtaga ctatacgtcg taggttcc 108

<210> 341

<211> 108

<212> DNA

<213>artificial sequence

<400> 341

gactacatgg gacatgtgag acatgagtga tttcagtcaa ataagtagaa ggcacataag 60

aaatttccaa aggcatgtct tcagtcatca ccatacgtcg taggttcc 108

<210> 342

<211> 108

<212> DNA

<213>artificial sequence

<400> 342

gactacatgg gacatgctgg atctgagttg gctccactgc cattgcggcc ccatcctcag 60

acaagccctc agcttgccta cgcactgcat tcatacgtcg taggttcc 108

<210> 343

<211> 108

<212> DNA

<213>artificial sequence

<400> 343

gactacatgg gacattgtac tgataccact gatgagaaat ttctagagcc ttttttcttc 60

tttgagacct caaatcctgt tcatggtgca gactacgtcg taggttcc 108

<210> 344

<211> 108

<212> DNA

<213>artificial sequence

<400> 344

gactacatgg gacatagatt caccccctgc tgaatttcag cctccagtgg ttcaagcaat 60

ttttgtatat ctgagttaaa ctgctccaat tcctacgtcg taggttcc 108

<210> 345

<211> 108

<212> DNA

<213>artificial sequence

<400> 345

gactacatgg gacatcgcgg tttgccatca agtttgctgc ttggtcacgt gtagagtcca 60

cctttgggcg tatgtcattc agttctgcct ttatacgtcg taggttcc 108

<210> 346

<211> 108

<212> DNA

<213>artificial sequence

<400> 346

gactacatgg gacatcggga ggtgacagct atccagttac tattcagaag actgagttta 60

tcttccacca acgtctcctt cttgcccaaa acttacgtcg taggttcc 108

<210> 347

<211> 108

<212> DNA

<213>artificial sequence

<400> 347

gactacatgg gacatgatct ctttgtcaat tccatatctg tagctgccag ccattctgtc 60

aagacattca tttcctttcg catcttacgg gactacgtcg taggttcc 108

<210> 348

<211> 108

<212> DNA

<213>artificial sequence

<400> 348

gactacatgg gacatagttc tttgggattt tccgtctgct ttttctgtac aatctgacgt 60

ccagtcttta tcaccatttc cacttcagac ttctacgtcg taggttcc 108

<210> 349

<211> 108

<212> DNA

<213>artificial sequence

<400> 349

gactacatgg gacatacgct gctcaaaatt ggctggtttc tggaataatc gaaacttcat 60

ggagacatct tgtaattttt tctgtaagga cagtacgtcg taggttcc 108

<210> 350

<211> 108

<212> DNA

<213>artificial sequence

<400> 350

gactacatgg gacattccac cttgtctgca atataagctg ccaactgctt gtcaatgaat 60

gtgagggact cctggattaa gtgtaaggat ttttacgtcg taggttcc 108

<210> 351

<211> 108

<212> DNA

<213>artificial sequence

<400> 351

gactacatgg gacatgcatg aaaatcaatc acaatatcac gtacattaag gaaataggct 60

ggaaaaaaaa ttagcaaaat catctcagtc atttacgtcg taggttcc 108

<210> 352

<211> 108

<212> DNA

<213>artificial sequence

<400> 352

gactacatgg gacatctccg ccatctgtta gggtctgtgc caatatgcga atctgatttg 60

ggttatcctc tgaatgtcgc atcaaatttt caatacgtcg taggttcc 108

<210> 353

<211> 108

<212> DNA

<213>artificial sequence

<400> 353

gactacatgg gacatcattc agcctagtgc agagccactg gtagttggtg gttagagttt 60

caagttcctt ttttaaggcc tcttgtgcta cagtacgtcg taggttcc 108

<210> 354

<211> 108

<212> DNA

<213>artificial sequence

<400> 354

gactacatgg gacataagat actcttcttc agcttgtgtc atccattcgt gcatctctga 60

tagatctttc tggaggctta cagttttctc caatacgtcg taggttcc 108

<210> 355

<211> 108

<212> DNA

<213>artificial sequence

<400> 355

gactacatgg gacatgcttc attctttatc ttctgcccac cttcattgac actgtttaga 60

ctgggctgaa ttgtctgaat atcactgact aaatacgtcg taggttcc 108

<210> 356

<211> 108

<212> DNA

<213>artificial sequence

<400> 356

gactacatgg gacataccag ctgggaggag agcttcttcc agcgtccctc aatttcttca 60

aattctgatt gatatttccg gctaatttca gagtacgtcg taggttcc 108

<210> 357

<211> 108

<212> DNA

<213>artificial sequence

<400> 357

gactacatgg gacatgttga ggtatggaga gtttggtttc tgactgctgg acccatgtcc 60

tgatggcact catggtctcc tgatagcgca ttgtacgtcg taggttcc 108

<210> 358

<211> 108

<212> DNA

<213>artificial sequence

<400> 358

gactacatgg gacatggaac atgggtcctt gtcctttctc tttcagggct atgctttgaa 60

tttttaatcg ttcaatttga ggttgaagat ctgtacgtcg taggttcc 108

<210> 359

<211> 108

<212> DNA

<213>artificial sequence

<400> 359

gactacatgg gacatctgat tatagaaagc gatgatgttg ttctgatact ccagccagtt 60

aagtctctca cttagcaact ggcagaattc gattacgtcg taggttcc 108

<210> 360

<211> 108

<212> DNA

<213>artificial sequence

<400> 360

gactacatgg gacatcagat aacaaagcac ggagtttaca agcagcacaa aatgagtaca 60

gatataaaaa ttaatgcata acctacattg acttacgtcg taggttcc 108

<210> 361

<211> 108

<212> DNA

<213>artificial sequence

<400> 361

gactacatgg gacatactgt tggagggttg ttggaatgtt gcaaagtatc cataaatctg 60

tatgtccact aagcgttgtc tataaacaca tgttacgtcg taggttcc 108

<210> 362

<211> 108

<212> DNA

<213>artificial sequence

<400> 362

gactacatgg gacattcagg ttgacaactg tggatcaagg cttttcattg cgttgtctgt 60

ctatgtatat gtaacccttt ggagtgatca atatacgtcg taggttcc 108

<210> 363

<211> 108

<212> DNA

<213>artificial sequence

<400> 363

gactacatgg gacataagaa gatcttgttt gagtgaatac agtttgccca tggattgctt 60

tttcttttct agatccgctt ttaaaacctg ttatacgtcg taggttcc 108

<210> 364

<211> 108

<212> DNA

<213>artificial sequence

<400> 364

gactacatgg gacattttga gaaggatgtc ttgtaaaaga acccagcggt cttctgtcca 60

tctacagatg tttgcccatc gatctcccaa tactacgtcg taggttcc 108

<210> 365

<211> 108

<212> DNA

<213>artificial sequence

<400> 365

gactacatgg gacatcacca tgtgagtgag agaattgacc ctgacttgtt cttgttctag 60

atcttcttga agcacctgaa agataaaatg ttttacgtcg taggttcc 108

<210> 366

<211> 108

<212> DNA

<213>artificial sequence

<400> 366

gactacatgg gacataaaaa caaataagga cttacttgct ttgtttttcc atgctagcta 60

ccctgaggca ttcccatctt gaatttagga gattacgtcg taggttcc 108

<210> 367

<211> 108

<212> DNA

<213>artificial sequence

<400> 367

gactacatgg gacattgagt atgaaactgg tctttcacca cttccacatc attagaaatc 60

tctccttgtg cttgcaatgt gtcctcagca gaatacgtcg taggttcc 108

<210> 368

<211> 108

<212> DNA

<213>artificial sequence

<400> 368

gactacatgg gacattcaga ggtggtgaca taagcagcct gtgtgtaggc atagctcttg 60

aatcgaggct taggggaaga agttctctca tattacgtcg taggttcc 108

<210> 369

<211> 108

<212> DNA

<213>artificial sequence

<400> 369

gactacatgg gacatcttca atgctcactt gttgaggcaa aacttggaag agtgatgtga 60

tgtacattaa gatggacttc ttatctggat aggtacgtcg taggttcc 108

<210> 370

<211> 108

<212> DNA

<213>artificial sequence

<400> 370

gactacatgg gacatacact attccagtca aataggtctg gcctaaaaca catacacata 60

cacacataca caaagacaaa tataaatcaa tcttacgtcg taggttcc 108

<210> 371

<211> 108

<212> DNA

<213>artificial sequence

<400> 371

gactacatgg gacatttacc tgccagtgga ggattatatt ccaaatcaaa ccaagagtca 60

gtttatgatt tccatctacg atgtcagtac ttctacgtcg taggttcc 108

<210> 372

<211> 108

<212> DNA

<213>artificial sequence

<400> 372

gactacatgg gacatcttaa aaataagtca cataccagtt tttgccctgt caggccttcg 60

aggaggtcta ggaggcgcct cccatcctgt aggtacgtcg taggttcc 108

<210> 373

<211> 108

<212> DNA

<213>artificial sequence

<400> 373

gactacatgg gacatgcccc tcaagtaact ggactgtaac ttgaatccag taaatgaact 60

acatgaaact gtcaccaaac taaagcaatt gattacgtcg taggttcc 108

<210> 374

<211> 108

<212> DNA

<213>artificial sequence

<400> 374

gactacatgg gacataagta acaaaccatt cttaccttag aaaattgtgc atttacccat 60

tttgtgaatg ttttcttttg aacatcttct ctttacgtcg taggttcc 108

<210> 375

<211> 108

<212> DNA

<213>artificial sequence

<400> 375

gactacatgg gacataaaat gacttaaact tcagcagcag gaaaagataa tctttactgt 60

ctgtctagtt ttagagcatc caataaacaa gaatacgtcg taggttcc 108

<210> 376

<211> 108

<212> DNA

<213>artificial sequence

<400> 376

gactacatgg gacatcttta ataatcctac cttccatgcc agctgttttt cctgtcactc 60

catcatgcca ataagttggc tgctgtaaca aattacgtcg taggttcc 108

<210> 377

<211> 108

<212> DNA

<213>artificial sequence

<400> 377

gactacatgg gacattacag tttctgtttt caatggttaa ttattcagct ggcaaagaga 60

aaggaaaaaa gggaaagctg ggagttggag ggatacgtcg taggttcc 108

<210> 378

<211> 108

<212> DNA

<213>artificial sequence

<400> 378

gactacatgg gacatgcacc atggacctgg agtgtggtaa caaagacaac aaagctttgc 60

ctggctggtg ggtgtgtgtg agtgtagggg agttacgtcg taggttcc 108

<210> 379

<211> 108

<212> DNA

<213>artificial sequence

<400> 379

gactacatgg gacatcagtg ctctgcttgg aggcaagtgg cacagagtgc ttacaaccag 60

agtgcctgaa tggggtctag ctgcagccag ttgtacgtcg taggttcc 108

<210> 380

<211> 108

<212> DNA

<213>artificial sequence

<400> 380

gactacatgg gacatcatgt tggggcctag cacccattta ggccctgggc atacggactt 60

cagtaagaca caatatggcg cttttggaca atgtacgtcg taggttcc 108

<210> 381

<211> 108

<212> DNA

<213>artificial sequence

<400> 381

gactacatgg gacatgaaaa agcatgagtc tttgtcttgt cctagaacaa tcaagagttg 60

atgtccctct gaggaactgg atgggttgaa cgctacgtcg taggttcc 108

<210> 382

<211> 108

<212> DNA

<213>artificial sequence

<400> 382

gactacatgg gacatctctg cccagattta ctgcctcaag acctctactc agcttccttc 60

tatacttagc tctactgata acaggagatg agttacgtcg taggttcc 108

<210> 383

<211> 108

<212> DNA

<213>artificial sequence

<400> 383

gactacatgg gacattcctc cagtggccct ttccaccctc ctgccatgtg tcccagatta 60

catctgccac caccccatat atatattcta tattacgtcg taggttcc 108

<210> 384

<211> 108

<212> DNA

<213>artificial sequence

<400> 384

gactacatgg gacatttcac aagacaaaag taaagctttt gaaggctcac catgtgtgtg 60

gactcgaaga ggattgttga tttcacatgt agatacgtcg taggttcc 108

<210> 385

<211> 108

<212> DNA

<213>artificial sequence

<400> 385

gactacatgg gacattgtca acacatattt atatatgcac attagtaggt ggcacctcca 60

agtctgtgtg tgatgggaga tatgcgagtg tgatacgtcg taggttcc 108

<210> 386

<211> 108

<212> DNA

<213>artificial sequence

<400> 386

gactacatgg gacatgctgt ccaaaacatg gagcttctgc aggcagaggg gccatgcctg 60

ggagcgcttc atcccattgg cctacagaga aagtacgtcg taggttcc 108

<210> 387

<211> 108

<212> DNA

<213>artificial sequence

<400> 387

gactacatgg gacattcaag aaggacaaca agggagaggg ttccccattt cttttgtatt 60

tgtttgtatt cttttctact tcttttctga tcttacgtcg taggttcc 108

<210> 388

<211> 108

<212> DNA

<213>artificial sequence

<400> 388

gactacatgg gacatgcggc aggtggttaa agtgagggaa ccctttttca gtttgggtgt 60

cacctctgat tattttgctg tcgctttccc tattacgtcg taggttcc 108

<210> 389

<211> 108

<212> DNA

<213>artificial sequence

<400> 389

gactacatgg gacatagtaa ccaggtttgt cttttagttg gagcaattgc tggctttgtg 60

gctgttagaa tcagtggaag cttttcaact atctacgtcg taggttcc 108

<210> 390

<211> 108

<212> DNA

<213>artificial sequence

<400> 390

gactacatgg gacattctct gatcagggtc agtcagaatc agcaatcctt ttccacagtg 60

ccctgcatag aaaactccaa tctttccatg atatacgtcg taggttcc 108

<210> 391

<211> 108

<212> DNA

<213>artificial sequence

<400> 391

gactacatgg gacatgttac cctggccctt gggtttcccc caccctcctc ccatcctttt 60

caggctcccg gaaagagaag acccccagtg aaatacgtcg taggttcc 108

<210> 392

<211> 108

<212> DNA

<213>artificial sequence

<400> 392

gactacatgg gacataagta gagtagatta aaaatggggt cttctattga attatttggg 60

tttcagaaag ttagagccat tcaagtggat gtatacgtcg taggttcc 108

<210> 393

<211> 108

<212> DNA

<213>artificial sequence

<400> 393

gactacatgg gacatagctg cctgttggct aggaaaagcc taggggattt gtgagaggcc 60

agaggctttt cataaatagg aattagggtt ctgtacgtcg taggttcc 108

<210> 394

<211> 108

<212> DNA

<213>artificial sequence

<400> 394

gactacatgg gacatgtgac atgggaatca ttttaccgca tcaccagggc tccgcctgcc 60

agcatgctgc ctttgatttg agaagcaaat acatacgtcg taggttcc 108

<210> 395

<211> 108

<212> DNA

<213>artificial sequence

<400> 395

gactacatgg gacatgcatc ccagtggcct ttccagagga aagtgtgaag gggcagtggg 60

agaaaaggtg agatgcaacc acagcaacac ccatacgtcg taggttcc 108

<210> 396

<211> 108

<212> DNA

<213>artificial sequence

<400> 396

gactacatgg gacataaata ttaagctagt gctcagaata ccagactatg gagtgggggg 60

cgttgcaggg cagaagttgc tttttgagat ctttacgtcg taggttcc 108

<210> 397

<211> 108

<212> DNA

<213>artificial sequence

<400> 397

gactacatgg gacatggagg acagctagat agtatagctc cttcaagggc agaggaagcc 60

aagaatagat attcctcccc tagcctggtc ccctacgtcg taggttcc 108

<210> 398

<211> 108

<212> DNA

<213>artificial sequence

<400> 398

gactacatgg gacatttgct ggtcagtgcc atgaaaatgt catctatgta aaccaagatg 60

cttgtacttg ggtagttctc tgtcagaaat ttttacgtcg taggttcc 108

<210> 399

<211> 108

<212> DNA

<213>artificial sequence

<400> 399

gactacatgg gacatagctt cagggctatg ttcaagaaag acaacttacc tcatctgctt 60

atgcagtgca tatgcttcaa tcaaagaatg cactacgtcg taggttcc 108

<210> 400

<211> 108

<212> DNA

<213>artificial sequence

<400> 400

gactacatgg gacatgcaca agttggttta gcaacacata ctcatattcc tactttccta 60

gaaattggtt ctcacccaca gataattata gggtacgtcg taggttcc 108

<210> 401

<211> 108

<212> DNA

<213>artificial sequence

<400> 401

gactacatgg gacatgggca ggccccactc caaccacccc cacccttcac cacccagccc 60

agcccatttt atctatctat agaaacctct aaatacgtcg taggttcc 108

<210> 402

<211> 108

<212> DNA

<213>artificial sequence

<400> 402

gactacatgg gacatttctc ctacagaata ggagtcttct tcatgctaca atgtgcactg 60

tgtacagaaa ctgtgaatag agaagtagcc atttacgtcg taggttcc 108

<210> 403

<211> 108

<212> DNA

<213>artificial sequence

<400> 403

gactacatgg gacatcctca acctcagtac gtccactgcg ttcaacatga actgtgtctc 60

cttctttccc caaacagaac aacttaagag atctacgtcg taggttcc 108

<210> 404

<211> 108

<212> DNA

<213>artificial sequence

<400> 404

gactacatgg gacatgcttg ggcagaatct agaagatgat tatgccccag ttggaccagg 60

gacagggctc ctgaaatttt tctggggaac caatacgtcg taggttcc 108

<210> 405

<211> 108

<212> DNA

<213>artificial sequence

<400> 405

gactacatgg gacatcattt aggtaggtct gaggaatcct gaagaacgtt ttagatcctt 60

agggattccg cggtttattc cgggggctgg gcttacgtcg taggttcc 108

<210> 406

<211> 108

<212> DNA

<213>artificial sequence

<400> 406

gactacatgg gacattgagg ctagctcatg gacattcttg aggcaagatc ttgtgcccgg 60

ccttaagagc tgtcagcctg cctaggcaga ggatacgtcg taggttcc 108

<210> 407

<211> 108

<212> DNA

<213>artificial sequence

<400> 407

gactacatgg gacatgaaat ctgctttttg ttttgcagag agcttggaga taattctggt 60

ggctgtgtgg agtatgtgtt ggaggtgagt tgctacgtcg taggttcc 108

<210> 408

<211> 108

<212> DNA

<213>artificial sequence

<400> 408

gactacatgg gacattggac tatgcttcat ctctctcctc cctaaaaaaa aatgcactcc 60

acagaaagct tgcatatgct tttagagcaa ttgtacgtcg taggttcc 108

<210> 409

<211> 108

<212> DNA

<213>artificial sequence

<400> 409

gactacatgg gacattaaaa gagctcttag aaaagttggt tttggtaagg ctggagattt 60

cttttttggc attgtttgaa gaagttctac ctatacgtcg taggttcc 108

<210> 410

<211> 108

<212> DNA

<213>artificial sequence

<400> 410

gactacatgg gacattccaa agtttacctg cagtatttaa gaagtggaga actaccagct 60

gagcaggcta ttccagtctg tagttcaccc gtttacgtcg taggttcc 108

<210> 411

<211> 108

<212> DNA

<213>artificial sequence

<400> 411

gactacatgg gacatatagt ccaaactttc tctgtgtctg gctagtattg aaaactagat 60

aaactgctcc aaaccaacat ggagtaaaga gcatacgtcg taggttcc 108

<210> 412

<211> 108

<212> DNA

<213>artificial sequence

<400> 412

gactacatgg gacatgttag catggcttca cactgtttgt gtagccaaga ggacagaatt 60

acatgaatga cagtgcccag agtgacagct gtatacgtcg taggttcc 108

<210> 413

<211> 108

<212> DNA

<213>artificial sequence

<400> 413

gactacatgg gacatgggca agggatgatt attagagagg gtgacaagga attaagtgtt 60

aggtccttac tcctgagtcc tttagaaaca ctgtacgtcg taggttcc 108

<210> 414

<211> 108

<212> DNA

<213>artificial sequence

<400> 414

gactacatgg gacatctcta taaacgactt ctcaattctc gcaaatagta gcagaaacct 60

tattaagaaa ggtatagtgt tcaaaatgta acctacgtcg taggttcc 108

<210> 415

<211> 108

<212> DNA

<213>artificial sequence

<400> 415

gactacatgg gacattgaag taattcagac agaagtgact tgtctgctct ctcccaggtc 60

acatgtctca gggaagagat ctgctaccca acctacgtcg taggttcc 108

<210> 416

<211> 108

<212> DNA

<213>artificial sequence

<400> 416

gactacatgg gacatctcgc catcagagaa ttgctttcac aagtgcaggg gtctctttaa 60

aatctctttg gaatactgtg cttttatttc tactacgtcg taggttcc 108

<210> 417

<211> 108

<212> DNA

<213>artificial sequence

<400> 417

gactacatgg gacattggaa gctaacaggg agcacaggaa aaagagagat ttgcgtatat 60

catgccatga ggaccatttg cagtgggtga gggtacgtcg taggttcc 108

<210> 418

<211> 108

<212> DNA

<213>artificial sequence

<400> 418

gactacatgg gacatgtgtt tgtccagagg tatcttttca gatcacctta tttatcttga 60

tgaatctgtt taatcgctca ccttttctct taatacgtcg taggttcc 108

<210> 419

<211> 108

<212> DNA

<213>artificial sequence

<400> 419

gactacatgg gacatgacga aggaggagcc gagtgtccat cttgcgcatc tggcctcccg 60

catctaggca catgtttaag atgcttacac agctacgtcg taggttcc 108

<210> 420

<211> 108

<212> DNA

<213>artificial sequence

<400> 420

gactacatgg gacatactag tgttggagcg caagttccca gtgaaaacct ttagaataaa 60

attggaccgc agatatcagg tatgggtata ggatacgtcg taggttcc 108

<210> 421

<211> 108

<212> DNA

<213>artificial sequence

<400> 421

gactacatgg gacatatggc tccgctgtgc gacccagcag ttagcggcag gagggacaga 60

gggatgatgt caactcgttt tgccagccga gcatacgtcg taggttcc 108

<210> 422

<211> 108

<212> DNA

<213>artificial sequence

<400> 422

gactacatgg gacataaggg taagcagggt tctagaaatt tgtgttatgt tttctcccca 60

ctgtatttat ttctttggtt agtggtgcaa gaatacgtcg taggttcc 108

<210> 423

<211> 108

<212> DNA

<213>artificial sequence

<400> 423

gactacatgg gacatcattc agtcagatct ctgttagcat ggccactccc aaacccccat 60

tcaaaatagg tgacctaccc cttttaaagt gggtacgtcg taggttcc 108

<210> 424

<211> 108

<212> DNA

<213>artificial sequence

<400> 424

gactacatgg gacatacttt acgaaccaac cttttaacag taactctagg agagaggata 60

tcaaaaattg gcagtgaaaa attatagata agctacgtcg taggttcc 108

<210> 425

<211> 108

<212> DNA

<213>artificial sequence

<400> 425

gactacatgg gacatagaac agaaataaat acaaataggt ggggcaagag aaaagaaaca 60

tgtgaaaagg cccctggatg gtttaagtta tattacgtcg taggttcc 108

<210> 426

<211> 108

<212> DNA

<213>artificial sequence

<400> 426

gactacatgg gacatgtact tctctttagt gatctcagag aatggtaaac cactttctgt 60

aatcataggg tccccttaaa aagaaagaaa gaatacgtcg taggttcc 108

<210> 427

<211> 108

<212> DNA

<213>artificial sequence

<400> 427

gactacatgg gacatataag aaagggttca gtgacagtgc actgagtccc cattaataat 60

aaaatgtatt taatttgaag ttaccctttg ttttacgtcg taggttcc 108

<210> 428

<211> 108

<212> DNA

<213>artificial sequence

<400> 428

gactacatgg gacatagaga acacatttgg ctctccctaa gagagtatgt tgcctcgtgg 60

tctcttgggt ttgcagtggg ttcttgatgc tgatacgtcg taggttcc 108

<210> 429

<211> 108

<212> DNA

<213>artificial sequence

<400> 429

gactacatgg gacatactga agtcaaccag gaatagaaga gtcctcacaa acaaaagatc 60

tcccacctga ggggcaagac ccctctctat ccatacgtcg taggttcc 108

<210> 430

<211> 108

<212> DNA

<213>artificial sequence

<400> 430

gactacatgg gacatccttc tgtctagact gtgtcaatgc attagtcgct acccctgagt 60

gcctggcttt gtctcccctg tgcatgtggc cagtacgtcg taggttcc 108

<210> 431

<211> 108

<212> DNA

<213>artificial sequence

<400> 431

gactacatgg gacattcctg acccatacga atccttttag taagggatgc ctgaacagcc 60

actattccag tgtctctggt tcaaagcagt gcctacgtcg taggttcc 108

<210> 432

<211> 108

<212> DNA

<213>artificial sequence

<400> 432

gactacatgg gacatagagt aagccttttt ttattttccg ttttcctaat tagaatctat 60

ttctgggtga attttttccc ctatttctca gcatacgtcg taggttcc 108

<210> 433

<211> 108

<212> DNA

<213>artificial sequence

<400> 433

gactacatgg gacattgtct cataatcagt gaatagtttc atcacatact gtccacagct 60

gcgcctactg tccctgaaaa agatttcaca cattacgtcg taggttcc 108

<210> 434

<211> 108

<212> DNA

<213>artificial sequence

<400> 434

gactacatgg gacatcctcc cccttaaaac aaagccactc ttacagaaag aagtagagta 60

catcacaccc agaatagaca tctcccaatg ttttacgtcg taggttcc 108

<210> 435

<211> 108

<212> DNA

<213>artificial sequence

<400> 435

gactacatgg gacattgtgt gagtgttttt tttgaaacac tgccatgttt taacgactgg 60

gaagctttga tacttttaaa acaaataaac tcttacgtcg taggttcc 108

<210> 436

<211> 108

<212> DNA

<213>artificial sequence

<400> 436

gactacatgg gacattggtg ttctcctctt ctttatagaa gagtaaattt ttttcttttt 60

ttcttttttt tgcagtagga gttgatatgg acttacgtcg taggttcc 108

<210> 437

<211> 108

<212> DNA

<213>artificial sequence

<400> 437

gactacatgg gacatgcaat gtggggatgg agttggaaga aacttctttg ttgtttgtcc 60

ttactttttt ttcagataat gttgtgaggc ccctacgtcg taggttcc 108

<210> 438

<211> 108

<212> DNA

<213>artificial sequence

<400> 438

gactacatgg gacattccca cagagaagac aggtgagagt ccactgctga cagatgtcca 60

tggaggtctc cttttccgcc aagatgcagg gcttacgtcg taggttcc 108

<210> 439

<211> 108

<212> DNA

<213>artificial sequence

<400> 439

gactacatgg gacataagat ggcatagtgg ctttctgtat tgccttttga tatgggaatg 60

tacgtatgac agtaggttta gaggcccaca ccttacgtcg taggttcc 108

<210> 440

<211> 108

<212> DNA

<213>artificial sequence

<400> 440

gactacatgg gacattccag agtataatag ttctaaagtc agttatgtaa tgtgcagaga 60

atgaaagaaa gaagattggt agagaaagaa gactacgtcg taggttcc 108

<210> 441

<211> 108

<212> DNA

<213>artificial sequence

<400> 441

gactacatgg gacattctca aggaagcggg atagaagaca gcaaaatcta attacacatc 60

aaaaagccag aaaagaaaaa aacagccaaa acctacgtcg taggttcc 108

<210> 442

<211> 108

<212> DNA

<213>artificial sequence

<400> 442

gactacatgg gacatcaaca gttctcaaga agtgcttaga ttggatgcaa aagatgaatt 60

cagttctaac taaattgagt tcactcactt atgtacgtcg taggttcc 108

<210> 443

<211> 108

<212> DNA

<213>artificial sequence

<400> 443

gactacatgg gacatggtat cctcagtttt ctccattgta ttatggtagg cctagtcacc 60

tgtggcaaag ggggaccagg agaagtttct acctacgtcg taggttcc 108

<210> 444

<211> 108

<212> DNA

<213>artificial sequence

<400> 444

gactacatgg gacataaact agataaactg ctccaaacca acatggagta aagagcatat 60

tcactggttt atttgcagta atttgcaatt tgttacgtcg taggttcc 108

<210> 445

<211> 108

<212> DNA

<213>artificial sequence

<400> 445

gactacatgg gacatgacag aattacatga atgacagtgc ccagagtgac agctgtatat 60

tgctcagagc ttttatttct tatacctaga atatacgtcg taggttcc 108

<210> 446

<211> 108

<212> DNA

<213>artificial sequence

<400> 446

gactacatgg gacatgcagg cacaccggta gaaaaactag aggggtctgg agttcctagg 60

tgaaccccag cagcctaacc ctgtcaagtc tgttacgtcg taggttcc 108

<210> 447

<211> 108

<212> DNA

<213>artificial sequence

<400> 447

gactacatgg gacatcagaa accttattaa gaaaggtata gtgttcaaaa tgtaacctgt 60

ctcctaccac tcttggctcc taccacatgt caatacgtcg taggttcc 108

<210> 448

<211> 108

<212> DNA

<213>artificial sequence

<400> 448

gactacatgg gacatctaaa catacacgca tcagtttata ggtcaaatat ctactgcaaa 60

ctcttcaccg ctgtaactta cttgctactt acatacgtcg taggttcc 108

<210> 449

<211> 108

<212> DNA

<213>artificial sequence

<400> 449

gactacatgg gacataccga aggcaacagc ggcggtagtg agcgacactg cgcaggattt 60

catggaaaca acaaatttcc aagtcccacg acgtacgtcg taggttcc 108

<210> 450

<211> 108

<212> DNA

<213>artificial sequence

<400> 450

gactacatgg gacattttga aagctttctc agtaaagcat ggtgccaggc accaagtggt 60

tccttataga ggagccagag ttaattttga tgatacgtcg taggttcc 108

<210> 451

<211> 108

<212> DNA

<213>artificial sequence

<400> 451

gactacatgg gacatactaa ttaaatgtct gtcatggata ctaaaatgta tatacggcct 60

ttgtgcctgg acaacagtac atgtgtcagc tgttacgtcg taggttcc 108

<210> 452

<211> 108

<212> DNA

<213>artificial sequence

<400> 452

gactacatgg gacatataga aaaatgtagg caaacagaaa caactgagtt gacttgacag 60

cagtgatgtt gaaaaccagc ctccagtccc tactacgtcg taggttcc 108

<210> 453

<211> 108

<212> DNA

<213>artificial sequence

<400> 453

gactacatgg gacattcctg tgtgggacat acttcctgtt tcgtctctgc aagacgcact 60

tactgtattg cccctgtggg ttacaggctt ccttacgtcg taggttcc 108

<210> 454

<211> 108

<212> DNA

<213>artificial sequence

<400> 454

gactacatgg gacatcccag gccagcagca tgccccagca aaggagggag ggagcagcac 60

aggcaggcaa agcttctaaa agtacaatgt ggctacgtcg taggttcc 108

<210> 455

<211> 108

<212> DNA

<213>artificial sequence

<400> 455

gactacatgg gacataggca gagagattat gcatctattg tatcttcatt ttccgtctgg 60

gaaaactcat ggcactaatg agctctagac atatacgtcg taggttcc 108

<210> 456

<211> 108

<212> DNA

<213>artificial sequence

<400> 456

gactacatgg gacattggta tcttccattg gttatttttg gaccatagct tttactcagt 60

atttcatttg ctgtctgcca acccacctga tgatacgtcg taggttcc 108

<210> 457

<211> 108

<212> DNA

<213>artificial sequence

<400> 457

gactacatgg gacatcaatt cccaaaagaa gactggtaat aacatcagaa accatgcttc 60

taagggaagt ctgcagaggg agagaagtag gagtacgtcg taggttcc 108

<210> 458

<211> 108

<212> DNA

<213>artificial sequence

<400> 458

gactacatgg gacataaagg ttggtctttc aagtggaaac caaacctgaa ggcatcatgt 60

catccaggta ttcaggaagt agataaaaga gagtacgtcg taggttcc 108

<210> 459

<211> 108

<212> DNA

<213>artificial sequence

<400> 459

gactacatgg gacattatca tattgaaaaa gctaccttat aggcataaca ggagtcactt 60

actttcactt caacacccct gccatttctc acctacgtcg taggttcc 108

<210> 460

<211> 108

<212> DNA

<213>artificial sequence

<400> 460

gactacatgg gacatttctc cccactgtat ttatttcttt ggttagtggt gcaagaaatt 60

ctgttttcct gtagcaaatt aataaagcgt tcatacgtcg taggttcc 108

<210> 461

<211> 108

<212> DNA

<213>artificial sequence

<400> 461

gactacatgg gacataaccc ccattcaaaa taggtgacct acccctttta aagtgggtta 60

ctttaagatt tctgtagttt aacttctggc aagtacgtcg taggttcc 108

<210> 462

<211> 108

<212> DNA

<213>artificial sequence

<400> 462

gactacatgg gacatcactg ggggacattg gggccagcag tgtctcattt ggtcgtttag 60

caagggtgca tattgccaag atgtggtacg cagtacgtcg taggttcc 108

<210> 463

<211> 108

<212> DNA

<213>artificial sequence

<400> 463

gactacatgg gacatgagag gatatcaaaa attggcagtg aaaaattata gataagcaaa 60

aagctccttc tgaggtccag gccaggagat agttacgtcg taggttcc 108

<210> 464

<211> 108

<212> DNA

<213>artificial sequence

<400> 464

gactacatgg gacataaaga aacatgtgaa aaggcccctg gatggtttaa gttatatttt 60

catcagtcat ccagttaaga gttaaagaat gagtacgtcg taggttcc 108

<210> 465

<211> 108

<212> DNA

<213>artificial sequence

<400> 465

gactacatgg gacattggct tcatagccac cttcctcaat cacaatctga aagtataaga 60

aacaatatgg atgaactgtg aacagactgg aaatacgtcg taggttcc 108

<210> 466

<211> 108

<212> DNA

<213>artificial sequence

<400> 466

gactacatgg gacatacttt ctgtaatcat agggtcccct taaaaagaaa gaaagaaaac 60

atgaaacaaa agaaacaagg acacatccca gaatacgtcg taggttcc 108

<210> 467

<211> 108

<212> DNA

<213>artificial sequence

<400> 467

gactacatgg gacatcgggt ccctgttgct tccgccgcgg ccccctggct gctgagagcc 60

agctgtttat gctccacttc ccggagccct tcctacgtcg taggttcc 108

<210> 468

<211> 108

<212> DNA

<213>artificial sequence

<400> 468

gactacatgg gacatattaa taataaaatg tatttaattt gaagttaccc tttgtttcag 60

agggtggagt agctgctgtt tacattgagc ccatacgtcg taggttcc 108

<210> 469

<211> 108

<212> DNA

<213>artificial sequence

<400> 469

gactacatgg gacatgcctc gtggtctctt gggtttgcag tgggttcttg atgctgaata 60

gtggtgtgac gtccctttgg gctacatcca acatacgtcg taggttcc 108

<210> 470

<211> 108

<212> DNA

<213>artificial sequence

<400> 470

gactacatgg gacatcgctg gcatacaaat cacctccctt tgcaaaacaa aaaacaaaaa 60

aaaaaaaaca ccggacacag cacagtacac caatacgtcg taggttcc 108

<210> 471

<211> 108

<212> DNA

<213>artificial sequence

<400> 471

gactacatgg gacatgctac atgaggttaa aggagagaag cagaagagca aatggggaaa 60

gcaagattcc ttctgtttac ggcttacatt gtctacgtcg taggttcc 108

<210> 472

<211> 108

<212> DNA

<213>artificial sequence

<400> 472

gactacatgg gacatggaca cttaccacta agccacgtag gagctcagga gaggtcctgg 60

tgagcaacgg cctaacaggc cagtgacacc ccatacgtcg taggttcc 108

<210> 473

<211> 108

<212> DNA

<213>artificial sequence

<400> 473

gactacatgg gacatctaaa cctaatgtct ttcctctttt tattcctctt ttaatttcta 60

ccctgattct tttgtcttag attcaaagaa ttatacgtcg taggttcc 108

<210> 474

<211> 108

<212> DNA

<213>artificial sequence

<400> 474

gactacatgg gacataaagg gattcaaatg taaatatgag tggcacagaa aacctgtttg 60

gagtccattt tatgcatctg ttaacatttt gaatacgtcg taggttcc 108

<210> 475

<211> 108

<212> DNA

<213>artificial sequence

<400> 475

gactacatgg gacattcgtg gcttccaaaa acgtatttta ggtgtctcat gttgctttta 60

tggtacaact tctctagagt tgaaaatatt aattacgtcg taggttcc 108

<210> 476

<211> 108

<212> DNA

<213>artificial sequence

<400> 476

gactacatgg gacatccaca gctgcgccta ctgtccctga aaaagatttc acacataggt 60

ctagcctttc tcagtctggt atgtattctt gagtacgtcg taggttcc 108

<210> 477

<211> 108

<212> DNA

<213>artificial sequence

<400> 477

gactacatgg gacataccaa aatcagggaa aattctatta tttccacatt tcaatttgaa 60

taaaacctgg aggtgaatca gatgaagacc aggtacgtcg taggttcc 108

<210> 478

<211> 108

<212> DNA

<213>artificial sequence

<400> 478

gactacatgg gacatgcatg gatagggtcc acttcacctg taaaatacat gaagatagat 60

gacatagaag gtgcttccaa atccattccc acatacgtcg taggttcc 108

<210> 479

<211> 108

<212> DNA

<213>artificial sequence

<400> 479

gactacatgg gacattcgtg agtggacact tttccattgt ctttggattt cattttggga 60

tagagagtgt gagcagaaat aaggtcaagt agctacgtcg taggttcc 108

<210> 480

<211> 108

<212> DNA

<213>artificial sequence

<400> 480

gactacatgg gacattcgct ggtgcaccct gcaagaagct ggtgccgtgg tgcctttccg 60

ggccctgcac actggtctga gcaccatgtt ttttacgtcg taggttcc 108

<210> 481

<211> 108

<212> DNA

<213>artificial sequence

<400> 481

gactacatgg gacataaagt acttcattcc agttatagaa atagttccac atttattgga 60

aaacttaact gatgaagaac tgaaaaagga ggctacgtcg taggttcc 108

<210> 482

<211> 108

<212> DNA

<213>artificial sequence

<400> 482

gactacatgg gacatgaggt tagtatgaaa gttagacagt tctgggatct tgaatgatct 60

ctacttaaca cgtaaataaa ttaagctatg agatacgtcg taggttcc 108

<210> 483

<211> 108

<212> DNA

<213>artificial sequence

<400> 483

gactacatgg gacatttatg agttagaatc agcaggagtt ggtaatttga tatttgagaa 60

agcggagcag gaaagagatg ttaagcctac atctacgtcg taggttcc 108

<210> 484

<211> 108

<212> DNA

<213>artificial sequence

<400> 484

gactacatgg gacatgagca tattcactgg tttatttgca gtaatttgca atttgtcagt 60

gtataagaca catgcagggt gaagtgtaca gagtacgtcg taggttcc 108

<210> 485

<211> 108

<212> DNA

<213>artificial sequence

<400> 485

gactacatgg gacatgcgat gataatctaa catgggctaa atcgtgtact tctgattgct 60

tgtgctattc agatttgatg atcgtggacg gagtacgtcg taggttcc 108

<210> 486

<211> 108

<212> DNA

<213>artificial sequence

<400> 486

gactacatgg gacatcactg ttagaacaca ttgggacatt aatgggatgg ttcccattgg 60

aacataatgg aattgaactg aaaaatcaaa ttgtacgtcg taggttcc 108

<210> 487

<211> 108

<212> DNA

<213>artificial sequence

<400> 487

gactacatgg gacatcagct gtctccccca cttctgtgtt gggctcaacc gaaaagaaag 60

atttaaatgt gaagggaagt ctggacctgg gcttacgtcg taggttcc 108

<210> 488

<211> 108

<212> DNA

<213>artificial sequence

<400> 488

gactacatgg gacattgggt gacaagaagt ctgttattag atgaaaccat gaattcaagc 60

aactcctgtt cctgaaatgg aaatcttgat gtctacgtcg taggttcc 108

<210> 489

<211> 108

<212> DNA

<213>artificial sequence

<400> 489

gactacatgg gacatcagcc ctataagaga aagaaatgag gaatttcact gttgctagtt 60

atttgcttat tgtaaactgg atggtggcct gattacgtcg taggttcc 108

<210> 490

<211> 108

<212> DNA

<213>artificial sequence

<400> 490

gactacatgg gacataaact ttgtatctca gtcttctctg taccaaatcg tgttaacttg 60

cccccactgt tattggggac agaataaaca ttatacgtcg taggttcc 108

<210> 491

<211> 108

<212> DNA

<213>artificial sequence

<400> 491

gactacatgg gacatcttaa gtcttgctcc aatgcaagat gtatctccat tcaacagtca 60

aggccagaaa gacttaagag gggacagatg gcatacgtcg taggttcc 108

<210> 492

<211> 108

<212> DNA

<213>artificial sequence

<400> 492

gactacatgg gacatacagc aggtgccatg ctaactgctg gcagaaggaa tcccgccttc 60

ttgcatgcag gcactcgccg ggcttgggaa agatacgtcg taggttcc 108

<210> 493

<211> 108

<212> DNA

<213>artificial sequence

<400> 493

gactacatgg gacattagga gtgcagccag gtgagggggc tagaaagaac agaccaaggt 60

gggaagcaga atttgtaaat tctctcggct acatacgtcg taggttcc 108

<210> 494

<211> 108

<212> DNA

<213>artificial sequence

<400> 494

gactacatgg gacatgagca cgccgtcgtg gcccggcagc cctggtctgc gggccagccc 60

tccgcaagtg acttttaaag actttctagc ctctacgtcg taggttcc 108

<210> 495

<211> 108

<212> DNA

<213>artificial sequence

<400> 495

gactacatgg gacatgtggg ttactttaag atttctgtag tttaacttct ggcaaggctt 60

ttaagaatca cagtattcaa gaaattttct acttacgtcg taggttcc 108

<210> 496

<211> 108

<212> DNA

<213>artificial sequence

<400> 496

gactacatgg gacattcgtt tagcaagggt gcatattgcc aagatgtggt acgcaggaga 60

gctgcatgcg aagaagttct tatcagcaaa caatacgtcg taggttcc 108

<210> 497

<211> 108

<212> DNA

<213>artificial sequence

<400> 497

gactacatgg gacattaagc aaaaagctcc ttctgaggtc caggccagga gatagtagga 60

tttaagaaac aaacaaacaa aaacaaccac aaatacgtcg taggttcc 108

<210> 498

<211> 108

<212> DNA

<213>artificial sequence

<400> 498

gactacatgg gacataagaa aacatgaaac aaaagaaaca aggacacatc ccagaatact 60

atttttagca tctttgggca ggaattagac aactacgtcg taggttcc 108

<210> 499

<211> 108

<212> DNA

<213>artificial sequence

<400> 499

gactacatgg gacattgttt cagagggtgg agtagctgct gtttacattg agcccaagat 60

acggttttaa cagtaacggc tactggggtc cattacgtcg taggttcc 108

<210> 500

<211> 108

<212> DNA

<213>artificial sequence

<400> 500

gactacatgg gacatggtcc tggtgagcaa cggcctaaca ggccagtgac accccattcc 60

agtgctgcag gtgctcatat gaccttcgcc ccgtacgtcg taggttcc 108

<210> 501

<211> 15

<212> DNA

<213>artificial sequence

<400> 501

gactacatgg gacat 15

<210> 502

<211> 15

<212> DNA

<213>artificial sequence

<400> 502

ggaacctacg acgta 15

<210> 503

<211> 50

<212> DNA

<213>artificial sequence

<400> 503

aatgatacgg cgaccaccga gatctacact ctttccctac acgacgctct 50

<210> 504

<211> 66

<212> DNA

<213>artificial sequence

<220>

<221> misc_feature

<222> (25)..(32)

<223> n is a, c, g, or t

<400> 504

caagcagaag acggcatacg agatnnnnnn nngtgactgg agttcagacg tgtgctcttc 60

cgatct 66

<210> 505

<211> 21

<212> DNA

<213>artificial sequence

<400> 505

aatgatacgg cgaccaccga g 21

<210> 506

<211> 21

<212> DNA

<213>artificial sequence

<400> 506

caagcagaag acggcatacg a 21

<210> 507

<211> 42

<212> DNA

<213>artificial sequence

<220>

<221> misc_feature

<222> (34)..(41)

<223> n is a, c, g, or t

<400> 507

acactctttc cctacacgac gctcttccga tctnnnnnnn nt 42

<210> 508

<211> 29

<212> DNA

<213>artificial sequence

<220>

<221> misc_feature

<222> (1)..(8)

<223> n is a, c, g, or t

<400> 508

nnnnnnnnag atcggaagag cacacgtct 29

Claims (20)

1. one kind is for detecting whether No. 13 chromosomes of fetus, No. 18 chromosomes, No. 21 chromosomes and sex chromosome to be measured are non- The kit of euploid, including probe groups；

The probe groups include 500 specific probes；Each specific probe includes DNA fragmentation 2；

The DNA fragmentation 2 is the nucleotide sequence of No. 13 chromosome, the nucleotide sequence of No. 18 chromosome, No. 21 chromosomes A part in the nucleotide sequence of nucleotide sequence, the nucleotide sequence of X chromosome or Y chromosome.

2. kit as described in claim 1, it is characterised in that: the DNA fragmentation 2 is the list of 60-140 nucleotide composition Ssdna molecule.

3. kit as claimed in claim 1 or 2, it is characterised in that: the nucleosides of the DNA fragmentation 2 of 500 specific probes Acid sequence successively as sequence table sequence 1 from 5 ' ends the 16-93 sequences 500 to sequence table 16- from 5 ' ends Shown in 93.

4. the kit as described in claims 1 to 3 is any, it is characterised in that: each specific probe further includes 1 He of DNA fragmentation DNA fragmentation 3；The DNA fragmentation 1 is located at 5 ' ends of the DNA fragmentation 2；The DNA fragmentation 3 is located at the 3 ' of the DNA fragmentation 2 End.

5. kit as claimed in claim 4, it is characterised in that: the DNA fragmentation 1 and the DNA fragmentation 3 are 10-20 The single strand dna of a nucleotide composition.

6. kit as claimed in claim 1 to 5, it is characterised in that: the kit further includes primer pair first；It is described Primer pair first is made of primer 1 and primer 2；The primer 1 includes the DNA fragmentation 1；The primer 2 includes the DNA fragmentation 3。

7. kit as claimed in claim 6, it is characterised in that: the primer 1 and/or the primer 2 have biotin mark Note.

8. kit as claimed in claim 1 to 5, it is characterised in that: the DNA fragmentation 1 and/or the DNA fragmentation 3 With biotin labeling.

9. kit as described in any of the claims 1 to 8, it is characterised in that: the kit further include connector 1, connector 2, Primer 3, primer 4, primer 5 and primer 6；

The connector 1 from 5 ' ends to 3 ' ends successively include DNA fragmentation a, DNA fragmentation b and DNA fragmentation c；The DNA fragmentation c 3 ' ends be T；

The connector 2 from 5 ' ends to 3 ' ends successively include DNA fragmentation d and DNA fragmentation e；

The DNA fragmentation b and the DNA fragmentation d are made of at random A, T, C and G；

3 ' end reverse complementals of the 5 ' ends of the DNA fragmentation e and the DNA fragmentation a；

The primer 5 includes the DNA fragmentation A；

The primer 3 from 5 ' ends to 3 ' ends successively include the DNA fragmentation A, DNA fragmentation B and DNA fragmentation C；The DNA 5 ' the ends of segment C and the DNA fragmentation a are completely the same；The DNA fragmentation B is made of at random A, T, C and G；

The primer 6 includes the DNA fragmentation D；

The primer 4 from 5 ' ends to 3 ' ends successively include the DNA fragmentation D, DNA fragmentation E and DNA fragmentation F；The DNA 3 ' the ends of segment F and the DNA fragmentation e are completely the same；The DNA fragmentation E is made of at random A, T, C and G.

10. kit as claimed in claim 9, it is characterised in that:

The length of the DNA fragmentation b and the DNA fragmentation d are 6-14bp；

The length of the DNA fragmentation A is 17-25bp；

The length of the DNA fragmentation B is 1-8bp；

The length of the DNA fragmentation D is 17-25bp；

The length of the DNA fragmentation E is 10-25bp.

11. the kit as described in claim 9 or 10, it is characterised in that: the primer 4 further includes Barcode flag sequence； The Barcode flag sequence is located at upstream, downstream or the centre of the DNA fragmentation E.

12. the kit as described in claim 9 to 11 is any, it is characterised in that: the connector 1, the primer 5 and described draw 3 ' ends of object 6 carry out thio-modification；5 ' ends of the connector 2 carry out phosphorylation modification.

13. the kit as described in claim 1 to 12 is any, it is characterised in that: the kit further includes recording judgement The carrier of standard first and judgment criteria second；

The judgment criteria first is to judge that the target autosome number of pregnant woman fetus to be measured is to increase, reduce or normal；Institute Stating target autosome is No. 13 chromosomes, No. 18 chromosomes or No. 21 chromosomes；

The judgment criteria first is as follows: first calculating pregnant woman's target autosome Z-score to be measured according to formula (1)；

ChrN_Z-score: pregnant woman's target autosome z-score to be measured；

%ChrN_sample: the reads ratio of pregnant woman's target autosome capture region to be measured；

Mean ChrN reference: the average value of target autosome capture region reads ratio in reference database；

S.D.%ChrN reference: the standard variance of target autosome capture region reads ratio in reference database；

Then it makes the following judgment: if target autosome Z-score >=3 of pregnant woman to be measured, the mesh of pregnant woman fetus to be measured Autosome number is marked to increase；If the target autosome Z-score < 3 of -3 < pregnant woman to be measured, pregnant woman fetus to be measured Target autosome number is normal；If the target chromosome Z-score of pregnant woman to be measured≤- 3, the target of pregnant woman fetus to be measured Autosome number is reduced；

If target autosome number increases or target autosome number is reduced, fetus target autosome is non-multiple Body；If target autosome number is normal, fetus target autosome is euploid；

The judgment criteria second is to judge that the sex chromosome number of pregnant woman fetus to be measured is to increase, reduce or normal；

The judgment criteria second is as follows:

(1) sex of foetus is first judged according to the reads ratio of pregnant woman's Y chromosome capture region to be measured: if pregnant woman's Y chromosome to be measured Capture region reads ratio > 0.001, then sex of foetus is male；If pregnant woman's Y chromosome capture region reads ratio to be measured ≤ 0.001, then sex of foetus is women；

(2) if fetus is women, the X chromosome Z-score of pregnant woman to be measured is first calculated according to formula (2)；

ChrX_Z-score: pregnant woman's X chromosome z-score to be measured；

%ChrN_sample: the reads ratio of pregnant woman's X chromosome capture region to be measured；

Mean ChrN reference: the average value of X chromosome capture region reads ratio in female's fetal data library；

S.D.%ChrN reference: the standard variance of X chromosome capture region reads ratio in female's fetal data library；

Then it makes the following judgment: if X chromosome Z-score >=3 of pregnant woman to be measured, the X chromosome of pregnant woman fetus to be measured Number increases；If the X chromosome Z-score < 3 of -3 < pregnant woman to be measured, the X chromosome number of pregnant woman fetus to be measured are normal； If the X chromosome Z-score of pregnant woman to be measured≤- 3, the X chromosome number of pregnant woman fetus to be measured is reduced；

When sex of foetus is women, if X chromosome number increases or X chromosome number is reduced, fetus X chromosome is non-whole Times body；If X chromosome number is normal, fetus X chromosome is euploid；

(3) if fetus is male, the X chromosome Z-score of pregnant woman to be measured is obtained first, in accordance with following steps (a)-(c):

(a) normal male database X chromosome and Y chromosome ratio mean value are calculated according to formula (3):

C: normal male database X chromosome and Y chromosome ratio mean value；

%ChrX_MDB: the average value of X chromosome capture region reads ratio in normal male database；

%ChrY_MDB: the average value of Y chromosome capture region reads ratio in normal male database；

(b) the reads ratio of converted rear pregnant woman's X chromosome capture region to be measured is calculated according to formula (4):

%ChrX is the reads ratio of converted rear pregnant woman's X chromosome capture region to be measured；

%ChrX_sampleFor the reads ratio of pregnant woman's X chromosome capture region to be measured；

C is normal male database X chromosome and Y chromosome ratio mean value；

%ChrY_sampleFor the reads ratio of pregnant woman's Y chromosome capture region to be measured；

(c) pregnant woman's X chromosome Z-score to be measured is calculated according to formula (5):

ChrX_Z-sco_re: pregnant woman's X chromosome z-score to be measured；

%ChrX: the reads ratio of pregnant woman's X chromosome capture region to be measured after converted；

Mean ChrXreference: the average value of X chromosome capture region reads ratio in female's fetal data library；

S.D.%ChrX reference: the standard variance of X chromosome capture region reads ratio in female's fetal data library；

Then it makes the following judgment: if X chromosome Z-score >=3 of pregnant woman to be measured and pregnant woman's Y chromosome capture region to be measured Give a birth out in reads ratio < normal pregnancies male fetus Y chromosome capture region reads ratio average value, then it is to be measured pregnant The X chromosome number of woman fetus increases, Y chromosome number is normal；If X chromosome Z-score >=3 of pregnant woman to be measured and to be measured It gives a birth out in pregnant woman Y chromosome capture region reads ratio >=normal pregnancies the Y chromosome capture region reads ratio of male fetus The average value of example, then the Y chromosome number increase of pregnant woman fetus to be measured, X chromosome number are normal；If -3 < pregnant woman's to be measured X chromosome Z-score < 3, then the X chromosome of pregnant woman fetus to be measured and Y chromosome number are normal；If the X of pregnant woman to be measured Chromosome Z-score≤- 3, then the X chromosome number of pregnant woman fetus to be measured is reduced, and Y chromosome number is normal；

When sex of foetus is male, if the increase of X chromosome number or X chromosome number are reduced, Y chromosome number is normal, Fetus X chromosome is aneuploid；If Y chromosome number increases, X chromosome number is normal, fetus Y chromosome is non-whole Times body；If X chromosome and Y chromosome number are normal, fetus X chromosome and Y chromosome are euploid；

The reference database by M healthy fetus of giving a birth healthy pregnant women data and common group of data of N healthy males At；M and N is 50 or more natural number；

Normal male database is made of the data of H healthy males；The natural number that H is 30 or more；

Female's fetal data library is made of the give birth to data of healthy pregnant women of female fetus of R status；The natural number that R is 30 or more；

Person under test is healthy pregnant women, healthy male or the healthy pregnant women for female fetus of giving a birth out of childbirth healthy fetus；It obtains to be measured The step of data of person, is as follows:

(K1) person under test's plasma DNA library is prepared；The connector for preparing person under test's plasma DNA library is claim 9 To connector 1 described in 12 and the connector 2；

(K2) after completing step (K1), by any probe groups in person under test's plasma DNA library and claim 1 to 8 Hybridized, obtains hybrid product；

(K3) after completing step (K2), target sequence is collected from the hybrid product, is obtained target sequence and is captured library；

(K4) the target sequence capture library obtained step (K3) carries out high-flux sequence, obtains sequencing initial data；

(K5) the sequencing initial data for obtaining step (K4) cuts off joint sequence, low quality base, then filtering is compared to ginseng Examine the corresponding position of genome hg19；

(K6) after completing step (K5), the repetitive sequence due to caused by PCR amplification preference is removed, the data of the person under test are obtained.

14. it is a kind of detect fetal chromosomal to be measured whether be aneuploid method, in turn include the following steps:

(1) pregnant woman blood plasma dissociative DNA to be measured library is prepared；

(2) after completing step (1), by any probe groups in the pregnant woman blood plasma dissociative DNA to be measured library and claim 1 to 8 Hybridized, obtains hybrid product；

(3) after completing step (2), target sequence is collected from the hybrid product, is obtained target sequence and is captured library；

(4) after completing step (3), target sequence capture library is sequenced, fetus to be measured is judged according to sequencing data Whether No. 13 chromosomes, No. 18 chromosomes, No. 21 chromosomes and sex chromosome are aneuploid.

15. method as claimed in claim 14, it is characterised in that: in the step (1), it is free to prepare pregnant woman blood plasma to be measured The connector of DNA library is connector 1 and the connector 2 described in claim 9 to 12.

16. method as claimed in claim 14, it is characterised in that: in the step (2), " by pregnant woman blood plasma dissociative DNA to be measured Library is hybridized with the probe groups, obtains hybrid product " the step of it is as follows:

(2-1) prepares enrichment system；The enrichment system include the pregnant woman blood plasma dissociative DNA to be measured library, the probe groups, Primer 3 described in claim 9 to 12 and the primer 4；

(2-2) takes the enrichment system, and 60-70 DEG C of hybridization is enriched with 16h or more, obtains hybrid product.

17. method as claimed in claim 14, it is characterised in that: in the step (3), " collect target sequence from hybrid product The step of column, acquisition target sequence capture library ", is as follows:

(3-a) collects the hybrid product with magnetic bead；

The product that (3-b) is collected using step (3-1) is template, using primer 5 described in claim 9 to 12 and the primer 6 The primer pair of composition carries out PCR amplification, and pcr amplification product is target sequence capture library.

18. method as claimed in claim 14, it is characterised in that: in the step (4), " judged according to sequencing data to be measured Whether fetal chromosomal is aneuploid " judgment criteria be claim 13 described in judgment criteria first and/or judgment criteria Second.

19. target sequence captures library, it is made of the target sequence that any the method for claim 14 to 18 obtains.

20. any probe groups in claim 1 to 8.