CN103820564B - The composite amplification reagent kit of 25 STRs - Google Patents
The composite amplification reagent kit of 25 STRs Download PDFInfo
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- CN103820564B CN103820564B CN201410091492.XA CN201410091492A CN103820564B CN 103820564 B CN103820564 B CN 103820564B CN 201410091492 A CN201410091492 A CN 201410091492A CN 103820564 B CN103820564 B CN 103820564B
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Abstract
The present invention relates to a kind of composite amplification system simultaneously analyzing multiple str locus seat, it is characterised in that: 25 locus of composite amplification: Amelogenin, vWA, D10S1248, D2S441, D21S11, D18S51, D5S818, D7S820, D13S317, D16S539, FGA, D2S1338, D22S1045, D1S1656, D19S433, D6S1043, D12S391, D8S1179, D3S1358, CSF1PO, Penta D, Penta E, TH01, TPOX and DYS391.The invention still further relates to analyze method and the test kit of DNA sample, and their purposes simultaneously.
Description
Technical field:
The present invention relates to biological technical field.Further, the present invention relates to detect in human genome that there is polymorphism
Genetic marker gene.The present invention be more particularly directed in an amplification system, expand multiple short series connection with polymerase chain reaction simultaneously
Duplicate loci.
Background technology:
STR (being called for short STR) also referred to as microsatellite or simple sequence repeats (SSR), be that a class is widely present
Strand of dna connection repetitive sequence in eukaryotic gene group, core sequence is 2-6 base recurring unit.Str locus seat quantity
Greatly, branch is extensive, accounts for 3% (Intemational Human Genome Sequencing of whole genome
Consortium, 2001), and polymorphism is high, and its polymorphism is derived mainly from core sequence number of repetition difference between individuality,
And this species diversity follows mendelian inheritance in genetic process.Therefore, STR augmentation detection technology is widely used in individuality
Identify, relationship is identified and population genetic study.
STR composite amplification technology is legal medical expert's individuality identification and the technical way of paternity test at present, all over the world
DNA laboratory be used widely (" Forensic DNA Typing " second edition, John Butler).On record
Part analysis is used widely, provides strong evidence for cracking of cases.Along with development, a lot of countries utilize this technology to set up
The DNA data base of offender and suspect, namely to the DNA data analysis of convict under detention and suspect and be entered into
In data base, it is simple to compare and the work such as investigation.
STR composite amplification technology in early days is capable of in a reaction system str locus seat of amplification about 10,
Along with the extensive of application and the increase of comparing, the information of 10 locus offers can not meet requirement, producer both domestic and external
The product of the more limited loci that exploitation makes new advances, the AmpFISTR Identifiler test kit of such as American AB I company and the U.S.
Promega company16 test kits are all that 15 str locus seats add sex determining gene.Domestic also have class
As product, the such as Ministry of Public Security two DNATyper15 can realize 15 locus amplifications (DNATyper15 genes simultaneously
The research of seat and selection, Ye Jian etc., " Forensic Science ", 2007 03).
But, increasingly wider as the application of identification of means recently as DNA, user's locus number to test kit
Amount, information content and proliferation time have had higher requirement.In terms of DNA database establishment, test kit compatibility is also carried
Go out new requirement.Having about 2,000,000 parts in the current DNA data base of China, China will enter one in terms of DNA database establishment
Step is accelerated, and the data in DNA data base in 2013 will there are about accounting in the data base of the developed countries such as Britain more than 10,000,000 parts
The data of total population 10% (Http:// www.forensic.gov.uk.Asplen,2004).Along with China DNA data base builds
If scale will constantly expand, the effect of comparing is more and more important.Comparing is built upon STR composite amplification analysis and obtains
Data basis on, need each STR test kit have on gene locus compatibility (forensic DNA database in Chian,
" Chinese law medical journal ", Jiang Xianhua, the 5th phase of volume 12 in 2006).
13 core gene seats that currently used test kit specifies with the CODIS standard of the U.S. mostly (vWA,
D21S11、D18S51、D5S818、D7S820、D13S317、D16S539、FGA、D8S1179、D3S1358、CSF1PO、THO1、
TPOX), based on, at this, basis simply selects to increase other locus, during also portioned product deletes 13 core sequences
Portion gene seat.But be as user to locus quantity, information content and the compatibility of test kit higher demand, each factory
Business is proposed to be had more site and meets the test kit in up-to-date CODIS site.Each manufacturer test kit locus information is shown in Table 1.
So, if using the STR test kit of different manufacturer, importing to the data in DNA data base has locus difference.This
Sample is comparison when, and not all data are all effective, and simply the data of homologous genes seating portion can be with comparison, and different portion
Divide and just cannot apply.Such as, Identifiler test kit andBetween 16 test kits, identical str locus seat is
13, other two str locus seats are not owing to difference is so acting on when comparing.Further, these 13 can comparison gene
Seat data are reliable when of being used for getting rid of, but the most inadequate in the case of assert.Need more gene
Seat data.So, owing to each STR test kit locus is different, result in the waste of DNA data base's part data resource, and have
Effect property is inadequate.
Along with STR detection kit is more and more extensive in the application of client unit, rapid amplifying is also that test kit performance carries
The aspect risen, faster amplification rate greatly reduces the waste of time cost, improves work efficiency.Traditional reagent
Box proliferation time was at about 3 to 4 hours, and our company's previous generation Goldeneye20A test kit proliferation time is also left at 2.5 hours
Right.If an amplified reaction is completed in realizing 1 hour, the performance of test kit can be greatly promoted, also can improve work efficiency.
Therefore, DNA identifies that field needs to have a reaction and expands more limited loci, is provided that more information amount, has
The most compatible and faster amplification rate STR composite amplification body.If able to realized primary first-order equation amplification 25 in one hour
Individual above locus, is possible not only to save reagent cost, it is also possible to improves work efficiency, save human cost etc..
Table 1: the gene locus information of each main flow manufacturer's test kit
Note: black font represents CODIS13 gene locus ,+represent the locus being included ,-represent base not to be covered
Because of seat
Summary of the invention:
The present invention be directed to the demand, initially set up the composite amplification system of 25 str locus seats of one-time detection, this
A little str locus seats contain the full gene seat that each manufacturer is used the most both at home and abroad.
Described locus is Amelogenin, vWA, D10S1248, D2S441, D21S11, D18S51, D5S818,
D7S820、D13S317、D16S539、FGA、D2S1338、D22S1045、D1S1656、D19S433、D6S1043、D12S391、
D8S1179, D3S1358, CSF1PO, Penta D, Penta E, TH01, TPOX and DYS391.
The amplification system of the present invention comprises primer mixture, reaction buffer and thermal starting Taq archaeal dna polymerase etc..
Specific primer is separately designed at the flank of its repetitive sequence first against above-mentioned 25 locus.Design of primers is adopted
Primer5 software, every primer annealing temperature is used to be close to or higher than 60 DEG C.Primer dimer, other interaction can not be produced
Or cross reaction, amplified production length is between 90-480bp.Every pair of primer is carried out amplification assay and optimizes, until obtaining clear
Clear single amplified band.Primer sequence see table 2.
Table 2 composite amplification system each locus primer sequence
According to expanding fragment length etc., said gene seat is divided into 4 groups, first group comprise D22S1045, TPOX,
D13S317, D7S820, D1S1656, Penta E and D10S1248, second group comprise D8S1179, D5S818, D19S433,
D16S539, CSF1PO and Penta D, the 3rd group comprises D3S1358, vWA, D2S1338, D18S51, D6S1043 and D2S441,
4th group comprises Amelogenin, TH01, D12S391, D21S11, FGA and DYS391.Often organize respectively by different fluorescein marks
Note, often among group, each locus amplifications product separates according to difference in length, and two locus can not have overlap.Respectively often group is drawn
Thing carries out composite amplification test.After determining the situations such as this group does not has non-specific amplification phenomenon, no cross reaction, adjust every pair of primer
Concentration, make each fragment peak equalization in group reach more than 40%.
By 4 groups of primers respectively with blue, green, yellow and red fluorescence element labelling.One chain of a labelling in every pair of primer,
It is marked at 5 ' ends of primer.Blue markings may select 5-FAM (CF), 6-FAM (6-CF 5(6)-Carboxyfluorescein) or close
The fluorescein molecule of spectrum, Green Marker may select HEX (chlordene-6-methyl fluorescein), JOE (6-carboxyl-4,5-bis-chloro-2,
7-dimethoxyfluorescein succinimide ester) or the fluorescein molecule of close spectrum, yellow flag may select TMR (4-methyl-
6-carboxy-rhodamine) or the fluorescein molecule of close spectrum, red-label may select ROX (carboxy-X-rhodamine) or phase dipped beam
The fluorescein molecule of spectrum.By 4 groups of 25 locus composite amplifications, adjust each locus primer concentration according to product peak height situation,
Each locus peak value entirety harmony is made to reach more than 30%.It is multiple that the primer mixture obtained may be used for above-mentioned 25 locus
Close amplification.
The pcr amplification reaction of the present invention can be carried out in certain buffer system.Buffer system includes: 50mM KCI,
10mM Tris-HC1 (PH8.3,25 DEG C), 2.0mM MgCl2, 0.1mg/ml BSA's (bovine serum albumin) and each 0.2mM
dNTP.DNTP is four kinds of deoxyribonucleotide (dATP, dTTP, dCTP, dGTP) equimolar mixture.
Taq archaeal dna polymerase needed for reaction is thermal starting archaeal dna polymerase, and antibody is closed and modified or all may be used of chemical modification
With.Each amplification system (25 μ 1) of the present invention needs the Taq archaeal dna polymerase of 2U to 4U.
Amplification system on various reaction heat circulating instruments (such as ABI9700, ABI9600, ABI2720, Bio-Rad
ICycler, Bio-Rad C1000 etc.) use following program can obtain preferable result: 96 DEG C are incubated 2 minutes；98 DEG C of guarantors
Temperature 2 seconds, 60 DEG C are incubated 40 seconds, and 68 DEG C are incubated 20 seconds, and this step runs 30 circulations；4-10 DEG C of insulation.
Template DNA in the present invention is human genome DNA.By various conventional methods, such as paramagnetic particle method, phenol chloroform method,
The template DNA that the methods such as resin purification (" Molecular Cloning: A Laboratory handbook " third edition, Cold Spring Harbor Publications) are extracted all can obtain
Preferably result.DNA can be prepared by with undertissue or cell: blood (ecchymosis), seminal fluid (seminal stain), skeleton, hair, saliva
(salivary stain), perspiration, amniotic fluid etc. containing fetal cell.DNA profiling amount preferably can obtain in the range of 0.25ng to 4ng
Preferably amplification, template amount is the lowest may cause the inspection of some gene locus not measure, and the highest meeting of template amount causes non-specific
Property amplified production produce.
According to the response procedures amplification template DNA specified in above-mentioned reaction buffer system, each locus can be obtained and mix
The amplified production closed.Due to the fact that and have employed fluorescently-labeled primer, amplified production is also with fluorescent marker, and labelling
Thing can send under laser excitation and can pass through sequenator (such as ABI377,310DNA sequencer) or genetic analyzer
The optical signal that (such as ABI3130,3100genetic analyzer) identifies, so amplified production can pass through sequenator or heredity
Carry out electrophoresis on the instruments such as analyser and detection is analyzed.
The when that sequenator or genetic analyzer detecting, amplified production and molecular weight internal standard (marker,
Intemal lane standard), Methanamide mix according to a certain percentage, enter in instrument capillary tube or gel and be separated by electrophoresis.
Molecular weight internal standard is made up of the fluorescent label DNA fragment of a plurality of known length, is used for calculating pcr amplification product fragment length,
Such that it is able to judge gene type and with allelic ladder comparison.
Data after electrophoresis can be analyzed in the data analysis software such as GeneMapper, GeneMarker, GeneScan,
Obtain str locus typing collection of illustrative plates and data.
The present invention relates to
(1) a kind of composite amplification system simultaneously analyzing multiple str locus seat, it is characterised in that: 25 bases of composite amplification
Because of seat: Amelogenin, vWA, D10S1248, D2S441, D21S11, D18S51, D5S818, D7S820, D13S317,
D16S539、FGA、D2S1338、D22S1045、D1S1656、D19S433、D6S1043、D12S391、D8S1179、D3S1358、
CSF1PO, Penta D, Penta E, TH01, TPOX and DYS391.
(2) composite amplification system of the present invention, wherein said locus is divided into following combination: first group is
D13S317, TPOX, D22S1045, D7S820, D1S1656, Penta E and D10S1248；Second group be D8S1179,
D5S818, D19S433, D16S539, CSF1PO and Penta D；3rd group be D3S1358, vWA, D2S1338, D18S51,
D6S1043 and D2S441；4th group is Amelogenin, TH01, D12S391, D21S11, FGA and DYS391, wherein, and amplification
Primer first group be SEQ ID NO:1-14；Second group is SEQ ID NO:15-26；3rd group is SEQ ID NO:27-38；
4th group is SEQ ID NO:39-50.
(3) according to the composite amplification system described in item 2, wherein in a composite amplification reaction system, expand 25 simultaneously
Locus.
(4) according to the composite amplification system described in item 2, wherein locus is expanded by the pair of primers being positioned at these locus both sides
Increase, wherein every pair of primer has 5 ' ends of a primer to carry out fluorescein labelling.
(5) according to the composite amplification system described in item 1, wherein composite amplification multiple str locus seat is to utilize polymerase chain
Formula reaction is carried out.
(6) according to the composite amplification system described in item 2, wherein this system comprises primer mixture.
(7) a kind of method simultaneously analyzing DNA sample, it is characterised in that apply the composite amplification system of any of the above-described to examine
Survey DNA.
(8) according to the method described in item 7, wherein DNA sample includes blood, ecchymosis, seminal fluid, seminal stain, skeleton, hair, saliva
In liquid, salivary stain, perspiration and amniotic fluid one or more..
(9) primer sequence for composite amplification system shown in SEQ ID NO:1-50 or the mixing of described primer sequence
Thing.
(10) a kind of test kit for analyzing multiple str locus seat simultaneously, described locus is Amelogenin, vWA,
D10S1248、D2S441、D21S11、D18S51、D5S818、D7S820、D13S317、D16S539、FGA、D2S1338、
D22S1045、D1S1656、D19S433、D6S1043、D12S391、D8S1179、D3S1358、CSF1PO、Penta D、Penta
E, TH01, TPOX and DYS391, this test kit comprises the primer shown in SEQ ID NO:1-50.
The present invention also provides for having selected from the primer sequence described in table 2, wherein one or more, preferably 1-15, excellent
Select 1-10, the modified sequence that preferably 1-5 nucleotide is replaced, deletes and/or adds and obtains.
The invention still further relates to the primer sequence mixture for composite amplification system shown in table 2.
The invention still further relates to above-mentioned primer sequence, test kit and/or composite amplification system be used for analyzing multiple str locus
The purposes of seat.
Use the present invention have only to one-time detection, operating time, detection time and detectable cost all reduce 50% or
More.The present invention is used to have the highest compatibility, the problem of data compatibility before using the present invention not worry.
Accompanying drawing explanation
Fig. 1-1 and Fig. 1-2 be DNA sample use the present invention amplification after analyze collection of illustrative plates two parts, the right of Fig. 1-1 with
There is overlap on the left side of Fig. 1-2, and both can be the collection of illustrative plates that a width is complete with split.
Fig. 2-1 and Fig. 2-2 uses the analysis chart after the Goldeneye20A test kit amplification of basic point cognition company for DNA sample
Two parts of spectrum, the right of Fig. 2-1 has overlapping with the left side of Fig. 2-2, and both can be the collection of illustrative plates that a width is complete with split.
Detailed description of the invention
The present invention is further illustrated below by the mode of embodiment.The mesh that the following examples are merely to illustrate that
, and and unrestricted the scope of the present invention.
25 gene locus of embodiment 1 composite amplification also analyze its genotype
Blood is contributed by volunteer.Template DNA is extracted by chelex-100 method by blood, separately adds legal medical expert's typing standard
Product 9948.Amplified reaction is carried out on ABI9700 thermal cycler, and electrophoresis and detection are carried out on ABI3100 genetic analyzer, number
GeneMapper ID v3.2 software is used according to analyzing.Sample uses the SinoFiler test kit of ABI company and basic point to recognize simultaneously
Knowing the Goldeneye20A test kit detection of company, operational approach is carried out according to test kit description, and result is as comparison.This
Bright reagent used and material such as allelic ladder (ladder) are the conventional material that those skilled in the art commonly use.
1.1.chelex-100 method extracts DNA (concrete grammar reference " Forensic DNA Protocol ", Humana
Press, 1998)
1) take 3 μ l and add the blood of anticoagulant in 500 μ l centrifuge tubes
2) vibration mixing chelex solution, makes chelex fully suspend, and often pipe adds the chelex-100 (100-of 195 μ l5%
200mesh, purchased from Bio-Rad company), add 5 μ l E.C. 3.4.21.64s (20mg/ml, purchased from Tian Gen biochemical technology company limited)
3) oscillation sample, after 56 DEG C of temperature are bathed 2 hours on constant-temperature metal bath, takes out sample and vibrates 2 minutes,
4) boiling 8-10 minute, 13000rpm is centrifuged 3 minutes
5) careful sucking-off about 150 μ l supernatant, is transferred in new pipe, and 10 μ l PCR reaction systems take 1 μ l as template
1.2. polymerase chain reaction (PCR) amplification
1) taking buffer, primer mixture, Taq enzyme, be made into mixed liquor according to following table, after vibration mixing, subpackage is anti-to PCR
Ying Guanzhong, often pipe 25 μ l, add template DNA
2) thermal cycler is set according to following reaction condition and increases instrument (ABI9700PCR instrument), PCR reaction tube is put into instrument
Device starts amplification gene fragment.96 DEG C are incubated 2 minutes；98 DEG C are incubated 2 seconds, and 60 DEG C are incubated 40 seconds, and 68 DEG C are incubated 20 seconds
Clock, runs 30 circulations；4 DEG C are persistently incubated, until taking out sample
1.3., after amplified reaction terminates, take out reaction tube, carry out electrophoresis and detection with ABI3100 genetic analyzer
1) take (0.5 μ l molecular weight internal standard+10 μ l deionized formamide) × (sample number) and be made into mixed liquor
2) subpackage, often pipe 10 μ l after mixing, then it is separately added into 1 μ l amplified production and allelic ladder (ladder), letter
Liquid is collected centrifuge tube tube bottom by short being centrifuged
3) 95 DEG C of degeneration of sample 4 minutes, then rapid cooled on ice 4 minutes, make the complete degeneration of DNA and keep degeneration shape
State
4) sample is put in the sample tray of Genetic Analyser, instrument parameter (sample introduction voltage 3kV, sample injection time are set
10 seconds), start electrophoresis detection
5) after about 40 minutes, electrophoresis terminates, and obtains collection of illustrative plates and typing knot by GeneMapper software analysis experimental data
Really (see Fig. 1-1, Fig. 1-2, Fig. 2-1, Fig. 2-2 and table 3)
3 two kinds of test kits of table expand the genotypic results of same sample
Invention effect:
Along with the increase of complex locus number, due to the impact of competition, the relative equilibrium of each locus controls difficulty and adds
Greatly, the present invention, by experiment repeatedly, has initially set up the composite amplification system of 25 str locus seats of one-time detection, these STR
Locus incorporates most gene locus that each manufacturer is used the most both at home and abroad.Utilize this amplification system permissible
Disposable 25 locus: Amelogenin of detection, vWA, D10S1248, D2S441, D21S11, D18S51, D5S818,
D7S820、D13S317、D16S539、FGA、D2S1338、D22S1045、D1S1656、D19S433、D6S1043、D12S391、
D8S1179, D3S1358, CSF1PO, Penta D, Penta E, TH01, TPOX and DYS391.
Utilizing this system can obtain 25 locus information with once-through operation, therefore this system has the highest individual knowledge
Not rate, is equivalent to the information summation simultaneously utilizing 2-3 other like products to obtain, no matter at PCR amplification and genetic analyzer
Detection, all provides cost savings and manpower, improves work efficiency.In amplification link, will be with first two or 3 kinds of test kits
Amplification is reduced to the amplification of a kind of test kit, has saved the reagent cost of more than 50%, and the time shortens 50%.Detection, uses
2 kinds or 3 kinds of test kits need sample detection respectively, use the present invention to have only to one-time detection, operating time, detection time and inspection
Test agent cost all reduces 50% or more.
The present invention is used to have the highest compatibility, the problem of data compatibility before using the present invention not worry.By
Contain the full gene seat of main product that is the most domestic and that use before in these 25 locus, there is good compatibility
Property, can not only existing total data in compatible China DNA data base at present, and new generation product is also had the highest holding concurrently
Capacitive.
It is understood that above description is only the example of the present invention, thus the model that the claims in the present invention are protected
Enclose and not merely limit with particular disclosed herein.The embodiment of any equivalent will be considered the present invention's
Within the scope of.It is true that according to description above, the present invention is carried out relevant modifications and variations for those skilled in the art
From the point of view of will be all possible, thus, this modifications and variations also will fall within the scope of appended claims of the invention.
Claims (4)
1. analyze a composite amplification method for multiple str locus seat, its diagnosis being not used in disease and treatment, its feature simultaneously
It is:
Expand 25 locus in a composite amplification reaction system simultaneously, 25 locus: Amelogenin of composite amplification,
vWA、D10S1248、D2S441、D21S11、D18S51、D5S818、D7S820、D13S317、D16S539、FGA、D2S1338、
D22S1045、D1S1656、D19S433、D6S1043、D12S391、D8S1179、D3S1358、CSF1PO、Penta D、Penta
E, TH01, TPOX and DYS391；
Described locus is divided into following four groups:
First group is D13S317, TPOX, D22S1045, D7S820, D1S1656, Penta E and D10S1248；
Second group is D8S1179, D5S818, D19S433, D16S539, CSF1PO and Penta D；
3rd group is D3S1358, vWA, D2S1338, D18S51, D6S1043 and D2S441；
4th group is Amelogenin, TH01, D12S391, D21S11, FGA and DYS391；
Specific primer is separately designed so that each sheet in locus group for the above-mentioned flank being grouped in locus repetitive sequence
Section peak equalization reaches more than 40%, and primer is divided into following combination:
First group of primer is shown in SEQ ID NO: 1-14；
Second group of primer is shown in SEQ ID NO: 15-26；
3rd group of primer is shown in SEQ ID NO: 27-38；
4th group of primer is shown in SEQ ID NO: 39-50；Often group primer is respectively by different fluorescein labellings；
Wherein locus is expanded by the pair of primers being positioned at these locus both sides, wherein has the 5 ' of a primer to hold in every pair of primer
Carry out fluorescein labelling；By above-mentioned four groups of locus composite amplifications, adjust each locus primer concentration according to product peak height situation,
Each locus peak value entirety harmony is made to reach more than 30%；
This composite amplification system is that 25 μ l amplification systems include the mixture of 5 μ l primer SEQ ID1-50,12.5 μ l reaction bufferings
The thermal starting Taq archaeal dna polymerase of liquid and 2U～4U；
Described reaction buffer includes: 50mM KCl, 10mM Tris-HCl, 2.0mM MgCl2, 0.1mg/ml bovine serum albumin
DNTP with 0.2mM.
Composite amplification method the most according to claim 1, it is characterised in that use following procedure to carry out:
1) template DNA is made；
2) polymerase chain reaction (PCR) amplification,
Every 25 μ l amplification systems are made into mixed liquor, and after mixing, subpackage is in PCR reaction tube, 2.5 μ l template DNAs is added PCR anti-
Ying Guanzhong；
Thermal cycler is set according to following reaction condition, PCR reaction tube is put into beginning amplification gene fragment in thermal cycler,
95 DEG C are incubated 11 minutes；94 DEG C are incubated 30 seconds, and 60 DEG C are incubated 60 seconds, and 70 DEG C are incubated 60 seconds, run 30 circulations；60℃
It is incubated 30 minutes；4 DEG C are persistently incubated until taking out sample；
3) electrophoresis, detection
Take 0.5 μ l molecular weight internal standard and 10 μ l deionized formamides, be multiplied by sample number, be made into mixed liquor；
1 μ l amplified production and ladder is added, the centrifugal bottom that liquid is collected centrifuge tube with often pipe 10 μ l mixed liquor；
95 DEG C of degeneration of sample 4 minutes, then rapid cooled on ice 4 minutes, make the complete degeneration of DNA and keep denatured state；
Sample is placed in Genetic Analyser beginning electrophoresis detection；
Electrophoresis terminates, and analyzes experimental data and obtains collection of illustrative plates and genotypic results.
3. the primer as shown in SEQ ID NO:1-50 for the composite amplification method described in claim 1 or 2 is mixed
Compound.
4. the composite amplification reagent of 25 STRs applying composite amplification method described in claim 1 or 2
Box, this test kit comprises the primer shown in SEQ ID NO:1-50, and described primer is for composite amplification locus:
Amelogenin、vWA、D10S1248、D2S441、D21S11、D18S51、D5S818、D7S820、D13S317、D16S539、
FGA、D2S1338、D22S1045、D1S1656、D19S433、D6S1043、D12S391、D8S1179、D3S1358、CSF1PO、
Penta D, Penta E, TH01, TPOX and DYS391.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410091492.XA CN103820564B (en) | 2014-03-13 | 2014-03-13 | The composite amplification reagent kit of 25 STRs |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410091492.XA CN103820564B (en) | 2014-03-13 | 2014-03-13 | The composite amplification reagent kit of 25 STRs |
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| CN106011229B (en) * | 2016-04-26 | 2019-08-27 | 深圳华大法医科技有限公司 | The composite amplification systems of 18 STR bit points for people, kit and application thereof |
| CN106906292A (en) * | 2017-03-29 | 2017-06-30 | 司法部司法鉴定科学技术研究所 | A kind of 22 STR composite amplification methods and its kit |
| CN116987798A (en) * | 2023-09-26 | 2023-11-03 | 深圳荻硕贝肯精准医学有限公司 | Primer combination, kit and method for detecting short tandem repeat sequence |
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| WO2012067901A1 (en) * | 2010-11-15 | 2012-05-24 | Life Technologies Corporation | Methods and kits for multiplex amplification of short tandem repeat loci |
| JP2014515926A (en) * | 2011-05-12 | 2014-07-07 | ネットバイオ・インコーポレーテッド | Methods and compositions for rapid multiplex amplification of STR loci |
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