CN108060212A - DNA typing identification kit - Google Patents

DNA typing identification kit Download PDF

Info

Publication number
CN108060212A
CN108060212A CN201711282858.1A CN201711282858A CN108060212A CN 108060212 A CN108060212 A CN 108060212A CN 201711282858 A CN201711282858 A CN 201711282858A CN 108060212 A CN108060212 A CN 108060212A
Authority
CN
China
Prior art keywords
artificial sequence
dna
sed
primer
locus
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN201711282858.1A
Other languages
Chinese (zh)
Other versions
CN108060212B (en
Inventor
郑焱
陈晓燕
许晓芝
谢龙旭
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
GUANGZHOU KAIPU PHARMACEUTICAL TECHNOLOGY Co.,Ltd.
HYBRIBIO Ltd.
Shanghai Kaipu medical laboratory Co.,Ltd.
Original Assignee
Guangzhou Kaipu Medical Technology Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Guangzhou Kaipu Medical Technology Co Ltd filed Critical Guangzhou Kaipu Medical Technology Co Ltd
Priority to CN201711282858.1A priority Critical patent/CN108060212B/en
Publication of CN108060212A publication Critical patent/CN108060212A/en
Application granted granted Critical
Publication of CN108060212B publication Critical patent/CN108060212B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/6858—Allele-specific amplification

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Immunology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The invention discloses a kind of DNA typing identification kit, the primer for being used for multiplex PCR including one group, sequence is SEQ ID No.1 52.DNA typing identification kit of the present invention expands 22 gene locis simultaneously in same reaction tube, can detect gender site and 21 autosome sites simultaneously.By expanding the DNA fragmentation of 22 gene locis simultaneously in same reaction tube, diagnosis efficiency and diagnostic accuracy are greatly improved, cost is reduced, shortens detection time.

Description

DNA typing identification kit
Technical field
The present invention relates to gene field, specifically by PCR amplification reagent, 22 pairs of STR identifications special primers, 4 pairs in primers F The universal primer and tetra- parts of fluorescent calibration object Matrix of sequenator that the 5' ends at end are modified with different fluorescence form, The polymorphism of short tandem repeat is detected by using gene amplification, using Sanger sequenators as technology platform, so as to The polymorphism at each str locus seat of human chromosomal is detected.
Background technology
Foreword
The PCR partings of STR (Short Tandem Repeat, abbreviation STR) locus be the personal identification of legal medical expert both at home and abroad at present and The main technology of paternity test and developing direction.The whole world has a countries and regions more than 120 to handle a case using DNA technique, has every year Ten hundreds of DNA evidences are used by court, it has also become the very important instrument such as individual identification and solution paternity test.China Forensic dna research was proceeded by from 1987, Material Evidence Identification Center, Ministry of Public Security, each province and city and institute of some areas punishment section etc. are single at present Position Successful utilization DNA typing technology, obtains very big achievement in actually handling a case.Therefore, STRs partings are considered as 2nd generation method The core of DNA fingerprint technology is cured, is current medical jurisprudence individual identification both at home and abroad and the major technique developing direction of paternity identification.
10% is tandem repetitive sequence in human genome DNA, is known as satellite DNA.Wherein repeated fragment forms for 2~7bp Be known as microsatellite, also known as short tandem repeat, the number of different human body genome satellite DNA recurring unit is variable , therefore form extremely complex allele fragment length polymorphism.It is generally believed that human gene is averaged, every 6~10kb is just There is 1 str locus seat, provide the abundant source of high information gene seat with paternity test for the personal identification of legal medical expert.In practice, Multiple STR, which are used in combination, can generate hundreds of millions of genotype combinations, and the frequency that each combination occurs in group is all It is very low, it can assert suspect in very high probability level.Therefore, the PCR partings of str locus seat are domestic and international legal medical expert's individuals Identification and the most important technology of paternity test.
In such a situa-tion, medical jurisprudence Relationship iden- tification, population genetic result are carried out using STRs typing methods Analysis, genome and gene diagnosis, it has also become the trend of development.Its advantage is, directly reads the segment of destination locations Information, the accuracy of detection is highest in current all methods；Common PCR amplification can obtain a large amount of products；Operation letter Single, speed is fast；Detection range is wide, and site all in section can be detected.
Up to the present, in the product of STR Classification Identifications, although easy to operate, speed to be fast, and detection range is wide, can Site all in section is detected, but easily leads to the disequilibrium of multiplexed PCR amplification, causes same fluorescence detection channel In the signal magnitude in each locus site differ.Not yet occur the product that can solve signal difference very well at present, therefore explore A kind of new STR bit point combination and PCR amplification mode have great importance.
The content of the invention
In order to solve the above technical problems, the present invention provides a kind of DNA typing identification kits, gene magnification can be based on With STRs typing method platforms, the STR segments of human chromosomal gene loci are expanded, are read using Sanger sequenators Each locus site, draws diagnosis after being compared with standard items.
The DNA typing identification kit of the present invention, the primer for being used for multiplex PCR including one group, sequence are SEQ ID No.1-52。
The sequence of SEQ ID No.1-52 is as follows：
1 primer information of table
Title Base number Sequence number Sequence
GLF 45 SED ID No.1 agcactggacgactcatcgatccctgggctctgtaaagaatagtg
WG1F 42 SED ID No.2 agcactggacgactcatcgattcaatacagacagacagacag
WG2F 41 SED ID No.3 agcactggacgactcatcgatatcctcattgacagaattgc
WG3F 42 SED ID No.4 agcactggacgactcatcgatcctgtcctagccttcttatag
WG4F 41 SED ID No.5 agcactggacgactcatcgatttgctagttctgtggtctta
WG5F 41 SED ID No.6 agcactggacgactcatcgattgttggtatagagcagtgtt
WG23F-re 46 SED ID No.7 cgtactcacagtccactgtcagcatgtgtttatttatacatatgtg
WG6F 41 SED ID No.8 cgtactcacagtccactgtcaggactctgacccatctaacg
WG7F 43 SED ID No.9 cgtactcacagtccactgtcaggttagatagagataggacaga
WG9F 40 SED ID No.10 cgtactcacagtccactgtcacaccgaagacccctcctgt
WG10F 43 SED ID No.11 cgtactcacagtccactgtcatctgagtccagtgactgccact
WG12F 42 SED ID No.12 gtgactagctccatgctacagtaatcagtgagccaattcctt
WG13F 47 SED ID No.13 gtgactagctccatgctacagtgcagtgatttctgatattttggtgc
WG14F 41 SED ID No.14 gtgactagctccatgctacagtaagtagctgctgagtgatt
WG15F 40 SED ID No.15 gtgactagctccatgctacagtaatagcagcaggcagatg
WG16F 42 SED ID No.16 gtgactagctccatgctacagtattccaatcatagccacagt
WG11F-re 46 SED ID No.17 gtgactagctccatgctacagtgcattgtactagtctcagggctaa
WG18F 44 SED ID No.18 ctgcatgctctactggatcaacgagccacttcccataataaatc
WG19F 40 SED ID No.19 ctgcatgctctactggatcaacgcacagaacaggcactta
WG20F 42 SED ID No.20 ctgcatgctctactggatcaacttgtgataggtacagtagca
WG21F 42 SED ID No.21 ctgcatgctctactggatcaacagcatcaaggtagttaggta
WG22F 45 SED ID No.22 ctgcatgctctactggatcaacgtggagtacagcaatcacaacaa
GLR 45 SED ID No.23 cgcatctgactgcatagcatcatcagagcttaaactgggaagctg
WG1R 41 SED ID No.24 cgcatctgactgcatagcatcgcctacagagtgattccatt
WG2R 41 SED ID No.25 cgcatctgactgcatagcatccaggctgactatggagttat
WG3R 39 SED ID No.26 cgcatctgactgcatagcatctgtgcttcagtctcctca
WG4R 40 SED ID No.27 cgcatctgactgcatagcatcgaggtggagattgaagtga
WG5R 40 SED ID No.28 cgcatctgactgcatagcatcgaggtggtggatgtgttaa
WG23R-re 46 SED ID No.29 ctcgtagctgactgactcagagtgtataacaaaattcctatgatgg
WG6R 39 SED ID No.30 ctcgtagctgactgactcagaagccataggcagcccaaa
WG7R 39 SED ID No.31 ctcgtagctgactgactcagatgacttggctgagatgtg
WG9R 39 SED ID No.32 ctcgtagctgactgactcagacccaccctactcccacct
WG10R 41 SED ID No.33 ctcgtagctgactgactcagattccaacctgagtctgccaa
WG12R 39 SED ID No.34 cctagcacagctctgatcacaataggaggtggatggatg
WG13R 43 SED ID No.35 cctagcacagctctgatcacagcctgggcaacagaataagatt
WG14R 41 SED ID No.36 cctagcacagctctgatcacacagggcataacattatccaa
WG15R 41 SED ID No.37 cctagcacagctctgatcacagcagatgatgtatcagagga
WG16R 39 SED ID No.38 cctagcacagctctgatcacaaactacacaacacgcctt
WG11R-re 43 SED ID No.39 cctagcacagctctgatcacaagtttaagacaaggctggggaa
WG18R 42 SED ID No.40 gatcgtcagtgagcactacatcaggatgggtggatcaataga
WG19R 40 SED ID No.41 gatcgtcagtgagcactacatctgaactcctcaggtccaa
WG20R 40 SED ID No.42 gatcgtcagtgagcactacatcggcaggtgatatggcatt
WG21R 42 SED ID No.43 gatcgtcagtgagcactacatcttcatcactgtatcgtatcc
WG22R 44 SED ID No.44 gatcgtcagtgagcactacatctagccagaccctgtctcaaaaa
SPY1F 21 SED ID No.45 agcactggacgactcatcgat
SP1R 21 SED ID No.46 cgcatctgactgcatagcatc
SPY2F 21 SED ID No.47 cgtactcacagtccactgtca
SP2R 21 SED ID No.48 ctcgtagctgactgactcaga
SPY3F 22 SED ID No.49 gtgactagctccatgctacagt
SP3R 21 SED ID No.50 cctagcacagctctgatcaca
SPY4F 22 SED ID No.51 ctgcatgctctactggatcaac
SP4R 22 SED ID No.52 gatcgtcagtgagcactacatc
More than primer combination can be used for simultaneously expand 22 locus, i.e. Amelogenin, D16S539, D7S820, D22S1045、Penta E、D1S1677、D17S1301、D13S317、vWA、TH01、CSF1PO、D14S1434、D19S433、 FGA、D20S482、D5S818、D3S1358、D6S1043、TPOX、D9S1122、D8S1179、D10S1435。
The above-mentioned kit of the present invention, the primer SPY1F, SPY2F, SPY3F, the 5 ' ends of SPY4F are respectively provided with 6- Tetra- kinds of fluorescent markers of FAM, VIC, NED, PET, be respectively (6-FAM)-AGCACTGGACGACTCATCGAT, (VIC)- CGTACTCACAGTCCACTGTCA、(NED)-GTGACTAGCTCCATGCTACAGT、(PET)- CTGCATGCTCTACTGGATCAAC。
The above-mentioned kit of the present invention, the DNA fragmentation to differ in size that the nucleotide not waited using its number is formed and right The difference of these segment institute mark fluorescents is come the method analyzed.
The kit of the present invention, can also include the reaction solution for multiplex PCR, and reaction solution is made of the following components：
Table 2
Reagent forms Per person-portion content
10 × PCR Buffer 4.5μl
25 mM dNTPs 0.5μl
10 μM of primer 1-44 2-20nM/ primer
10 μM of primer 45-52 200nM/ primer
Archaeal dna polymerase 5U/ μ l 1.4μl
Add dd H2O is extremely 28μl
The present invention above-mentioned kit after product pretreatment, carries out Capillary Electrophoresis, big to different fragments using fluorescence detector Small product is identified and distinguishes.
The present invention, by largely testing adjustment, enables sample same anti-in identical conditions on the basis of design is rational It answers and the amplification of multidigit point is carried out at the same time in liquid, and based on gene magnification and STRs typing method platforms, by human chromosomal gene position The STR segments of point carry out Capillary Electrophoresis, each locus site are read using Sanger sequenators, after being compared with standard items Draw expert's conclusion.
The DNA typing identification kit of the present invention expands 22 gene locis simultaneously in same reaction tube, can be simultaneously Detect gender site and 21 autosome sites.By the DNA for expanding 22 gene locis simultaneously in same reaction tube Segment is greatly improved diagnosis efficiency and diagnostic accuracy, reduces cost, shortens detection time.
Description of the drawings
It is the explanation of attached drawing below, in order to understand the purpose of foregoing invention and specific features.
Fig. 1 be the invention detects that each locus specific distributing position and carry out Single tube amplification after each locus band Picture, each locus contain a pair of alleles.
Fig. 2 .1 are that 6 pairs of special primers without fluorescent decoration combine a pair of of universal primer, wherein a universal primer That is the 5' ends of SPY1F carry out 6-FAM modifications, each parameter are adjusted by testing, so as to fulfill the detection with fluorescence of 6 locus As a result picture.
Fig. 2 .2 are that 5 pairs of special primers without fluorescent decoration combine a pair of of universal primer, wherein a universal primer That is the 5' ends of SPY2F carry out VIC modifications, each parameter are adjusted by testing, so as to fulfill detection knot of 5 locus with fluorescence Fruit picture.
Fig. 2 .3 are that 6 pairs of special primers without fluorescent decoration combine a pair of of universal primer, wherein a universal primer That is the 5' ends of SPY3F carry out NED modifications, each parameter are adjusted by testing, so as to fulfill detection knot of 6 locus with fluorescence Fruit picture.
Fig. 2 .4 are that 5 pairs of special primers without fluorescent decoration combine a pair of of universal primer, wherein a universal primer That is the 5' ends of SPY4F carry out PET modifications, each parameter are adjusted by testing, so as to fulfill detection knot of 5 locus with fluorescence Fruit picture.
Fig. 3 is the collection of illustrative plates of the fluorescent calibration object Matrix of sequenator.
Specific embodiment
1. it is determined for expanding the design of the primer of DNA fragmentation and PCR reaction systems in measuring samples
1-1 design of primers
Sequence data according to obtained each locus is searched devises 44 common special primers, 8 universal primers wherein 4 With fluorescent marker, 52 primer combinations are placed in same reaction tube and carry out multiplex PCR, while 22 locus of amplification Segment.
1-2 multi-PRC reaction systems determine
By substantial amounts of Experimental comparison, 10 × PCR Buffer concentration is controlled（Contain Mg in 10 × PCR Buffer2+）、 Primer concentration and dNTPs concentration can accomplish the high efficiency amplification simultaneously of 22 locus, and specificity is good.It finally determines optimal PCR reaction systems are shown in Table 3：28 μ l of PCR reaction solution, 2 μ l of DNA profiling amount, total reaction volume are 30 μ l.
3 DNA typing of table identifies multi-PRC reaction system
Reagent forms Per person-portion content
10 × PCR Buffer 4.5μl
25 mM dNTPs 0.5μl
10 μM of primer 1-44 2-20nM/ primer
10 μM of primer 45-52 200nM/ primer
Archaeal dna polymerase 5U/ μ l 1.4μl
DNA profiling 2μl
Add dd H2O is extremely 30μl
1-3 multiplexed PCR amplification programs determine
It compares and optimizes by many experiments, control annealing temperature and annealing time that can accomplish that specificity is good, amplification efficiency is high, most Definite optimal amplification program is eventually：95 °C of thermal starting 15min；（94 °C of 45 sec of denaturation, 54 °C of annealing 10min, 72 °C of extensions 50 sec）, totally 2 cycle；（94 °C of 45 sec of denaturation, 56 °C of annealing 45sec, 72 °C of 50 sec of extension）, totally 28 cycle；Most Latter 72 °C of extension 10min of step.
It expands 22 locus simultaneously in the same reaction tube of identical conditions, reduces the demand of PCR instrument, reduce behaviour Make step, reduce cost, 22 gene locis can just be detected simultaneously by being once loaded, and improve identification accuracy rate.
The upper electromechanical swimming of 1-4 and atlas analysis
Capillary electrophoresis analysis is carried out to PCR product using 3500xL sequenators, and with G5-Matrix Standard to instrument Fluorescent calibration is carried out, while has write corresponding Panels, bins files, and with software GeneMapper ID version 3.0 carry out interpretation of result.
Sequence table
<110>Guangzhou Kai Pu Pharmaceutical Technology Co., Ltd
<120>DNA typing identification kit
<160> 52
<170> SIPOSequenceListing 1.0
<210> 1
<211> 45
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 1
agcactggac gactcatcga tccctgggct ctgtaaagaa tagtg 45
<210> 2
<211> 42
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 2
agcactggac gactcatcga ttcaatacag acagacagac ag 42
<210> 3
<211> 41
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 3
agcactggac gactcatcga tatcctcatt gacagaattg c 41
<210> 4
<211> 42
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 4
agcactggac gactcatcga tcctgtccta gccttcttat ag 42
<210> 5
<211> 41
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 5
agcactggac gactcatcga tttgctagtt ctgtggtctt a 41
<210> 6
<211> 41
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 6
agcactggac gactcatcga ttgttggtat agagcagtgt t 41
<210> 7
<211> 46
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 7
cgtactcaca gtccactgtc agcatgtgtt tatttataca tatgtg 46
<210> 8
<211> 41
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 8
cgtactcaca gtccactgtc aggactctga cccatctaac g 41
<210> 9
<211> 43
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 9
cgtactcaca gtccactgtc aggttagata gagataggac aga 43
<210> 10
<211> 40
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 10
cgtactcaca gtccactgtc acaccgaaga cccctcctgt 40
<210> 11
<211> 43
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 11
cgtactcaca gtccactgtc atctgagtcc agtgactgcc act 43
<210> 12
<211> 42
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 12
gtgactagct ccatgctaca gtaatcagtg agccaattcc tt 42
<210> 13
<211> 47
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 13
gtgactagct ccatgctaca gtgcagtgat ttctgatatt ttggtgc 47
<210> 14
<211> 41
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 14
gtgactagct ccatgctaca gtaagtagct gctgagtgat t 41
<210> 15
<211> 40
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 15
gtgactagct ccatgctaca gtaatagcag caggcagatg 40
<210> 16
<211> 42
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 16
gtgactagct ccatgctaca gtattccaat catagccaca gt 42
<210> 17
<211> 46
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 17
gtgactagct ccatgctaca gtgcattgta ctagtctcag ggctaa 46
<210> 18
<211> 44
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 18
ctgcatgctc tactggatca acgagccact tcccataata aatc 44
<210> 19
<211> 40
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 19
ctgcatgctc tactggatca acgcacagaa caggcactta 40
<210> 20
<211> 42
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 20
ctgcatgctc tactggatca acttgtgata ggtacagtag ca 42
<210> 21
<211> 42
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 21
ctgcatgctc tactggatca acagcatcaa ggtagttagg ta 42
<210> 22
<211> 45
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 22
ctgcatgctc tactggatca acgtggagta cagcaatcac aacaa 45
<210> 23
<211> 45
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 23
cgcatctgac tgcatagcat catcagagct taaactggga agctg 45
<210> 24
<211> 41
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 24
cgcatctgac tgcatagcat cgcctacaga gtgattccat t 41
<210> 25
<211> 41
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 25
cgcatctgac tgcatagcat ccaggctgac tatggagtta t 41
<210> 26
<211> 39
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 26
cgcatctgac tgcatagcat ctgtgcttca gtctcctca 39
<210> 27
<211> 40
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 27
cgcatctgac tgcatagcat cgaggtggag attgaagtga 40
<210> 28
<211> 40
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 28
cgcatctgac tgcatagcat cgaggtggtg gatgtgttaa 40
<210> 29
<211> 46
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 29
ctcgtagctg actgactcag agtgtataac aaaattccta tgatgg 46
<210> 30
<211> 39
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 30
ctcgtagctg actgactcag aagccatagg cagcccaaa 39
<210> 31
<211> 39
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 31
ctcgtagctg actgactcag atgacttggc tgagatgtg 39
<210> 32
<211> 39
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 32
ctcgtagctg actgactcag acccacccta ctcccacct 39
<210> 33
<211> 41
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 33
ctcgtagctg actgactcag attccaacct gagtctgcca a 41
<210> 34
<211> 39
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 34
cctagcacag ctctgatcac aataggaggt ggatggatg 39
<210> 35
<211> 43
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 35
cctagcacag ctctgatcac agcctgggca acagaataag att 43
<210> 36
<211> 41
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 36
cctagcacag ctctgatcac acagggcata acattatcca a 41
<210> 37
<211> 41
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 37
cctagcacag ctctgatcac agcagatgat gtatcagagg a 41
<210> 38
<211> 39
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 38
cctagcacag ctctgatcac aaactacaca acacgcctt 39
<210> 39
<211> 43
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 39
cctagcacag ctctgatcac aagtttaaga caaggctggg gaa 43
<210> 40
<211> 42
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 40
gatcgtcagt gagcactaca tcaggatggg tggatcaata ga 42
<210> 41
<211> 40
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 41
gatcgtcagt gagcactaca tctgaactcc tcaggtccaa 40
<210> 42
<211> 40
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 42
gatcgtcagt gagcactaca tcggcaggtg atatggcatt 40
<210> 43
<211> 42
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 43
gatcgtcagt gagcactaca tcttcatcac tgtatcgtat cc 42
<210> 44
<211> 44
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 44
gatcgtcagt gagcactaca tctagccaga ccctgtctca aaaa 44
<210> 45
<211> 21
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 45
agcactggac gactcatcga t 21
<210> 46
<211> 21
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 46
cgcatctgac tgcatagcat c 21
<210> 47
<211> 21
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 47
cgtactcaca gtccactgtc a 21
<210> 48
<211> 21
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 48
ctcgtagctg actgactcag a 21
<210> 49
<211> 22
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 49
gtgactagct ccatgctaca gt 22
<210> 50
<211> 21
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 50
cctagcacag ctctgatcac a 21
<210> 51
<211> 22
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 51
ctgcatgctc tactggatca ac 22
<210> 52
<211> 22
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 52
gatcgtcagt gagcactaca tc 22

Claims (3)

1.DNA Classification Identification kits, which is characterized in that the primer for being used for multiplex PCR including one group, sequence are SEQ ID No.1-52。
2. kit according to claim 1, which is characterized in that the reaction solution including being used for multiplex PCR, reaction solution press everyone Part content is made of the following components：
3. kit according to claim 1, which is characterized in that 5 ' ends of primer SED ID No.45,47,49,51, There are tetra- kinds of fluorescent markers of 6-FAM, VIC, NED, PET respectively.
CN201711282858.1A 2017-12-07 2017-12-07 DNA typing identification kit Active CN108060212B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201711282858.1A CN108060212B (en) 2017-12-07 2017-12-07 DNA typing identification kit

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201711282858.1A CN108060212B (en) 2017-12-07 2017-12-07 DNA typing identification kit

Publications (2)

Publication Number Publication Date
CN108060212A true CN108060212A (en) 2018-05-22
CN108060212B CN108060212B (en) 2019-06-04

Family

ID=62135405

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201711282858.1A Active CN108060212B (en) 2017-12-07 2017-12-07 DNA typing identification kit

Country Status (1)

Country Link
CN (1) CN108060212B (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116949223A (en) * 2023-09-19 2023-10-27 广东凯普生物科技股份有限公司 Hepatitis B virus drug administration guidance system and application thereof

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103789413A (en) * 2014-01-06 2014-05-14 基点认知技术(北京)有限公司 Multiplex amplification kit of 18 short tandem repeats (STR)
CN103834732A (en) * 2014-02-21 2014-06-04 苏州阅微基因技术有限公司 composite amplification system of 23 short tandem repeat sequences and a kit
CN105385763A (en) * 2015-12-11 2016-03-09 无锡中德美联生物技术有限公司 Fluorescence labeled multiplex amplification kit for simultaneously analyzing 24 loci of human genome DNA, and application thereof
CN105969905A (en) * 2016-07-27 2016-09-28 中国科学院北京基因组研究所 Detection kit for 38 STR allelic genes based on high-throughput sequencing
CN106048053A (en) * 2016-08-03 2016-10-26 中国科学院北京基因组研究所 Detection kit for 38 SRT alleles based on high-throughput sequencing
CN106086210A (en) * 2016-08-03 2016-11-09 中国科学院北京基因组研究所 40 allelic detection kit of STR based on high-flux sequence
CN106222258A (en) * 2016-07-27 2016-12-14 中国科学院北京基因组研究所 35 allelic detection kit of STR based on high-flux sequence
CN107012225A (en) * 2017-04-20 2017-08-04 司法部司法鉴定科学技术研究所 A kind of detection kit and detection method of the str locus seat based on high-flux sequence

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103789413A (en) * 2014-01-06 2014-05-14 基点认知技术(北京)有限公司 Multiplex amplification kit of 18 short tandem repeats (STR)
CN103834732A (en) * 2014-02-21 2014-06-04 苏州阅微基因技术有限公司 composite amplification system of 23 short tandem repeat sequences and a kit
CN105385763A (en) * 2015-12-11 2016-03-09 无锡中德美联生物技术有限公司 Fluorescence labeled multiplex amplification kit for simultaneously analyzing 24 loci of human genome DNA, and application thereof
CN105969905A (en) * 2016-07-27 2016-09-28 中国科学院北京基因组研究所 Detection kit for 38 STR allelic genes based on high-throughput sequencing
CN106222258A (en) * 2016-07-27 2016-12-14 中国科学院北京基因组研究所 35 allelic detection kit of STR based on high-flux sequence
CN106048053A (en) * 2016-08-03 2016-10-26 中国科学院北京基因组研究所 Detection kit for 38 SRT alleles based on high-throughput sequencing
CN106086210A (en) * 2016-08-03 2016-11-09 中国科学院北京基因组研究所 40 allelic detection kit of STR based on high-flux sequence
CN107012225A (en) * 2017-04-20 2017-08-04 司法部司法鉴定科学技术研究所 A kind of detection kit and detection method of the str locus seat based on high-flux sequence

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116949223A (en) * 2023-09-19 2023-10-27 广东凯普生物科技股份有限公司 Hepatitis B virus drug administration guidance system and application thereof
CN116949223B (en) * 2023-09-19 2023-12-29 广东凯普生物科技股份有限公司 Hepatitis B virus drug administration guidance system and application thereof

Also Published As

Publication number Publication date
CN108060212B (en) 2019-06-04

Similar Documents

Publication Publication Date Title
CN1896284B (en) Method for identifying allelic gene type
CN105695572B (en) Method for developing molecular markers in large scale and efficiently based on Indel and SSR site technology
US11789906B2 (en) Systems and methods for genomic manipulations and analysis
Tamaki et al. Human tandem repeat sequences in forensic DNA typing
He et al. Forensic features and phylogenetic analyses of Sichuan Han population via 23 autosomal STR loci included in the Huaxia Platinum System
US8315816B2 (en) Methods of genetic analysis involving the amplification of complementary duplicons
CN110863056A (en) Method, reagent and application for accurately typing human DNA
CN112592981B (en) Primer group, kit and method for DNA archive construction
CN104031989B (en) The test kit of the composite amplification of a kind of human gene group DNA 26 locus
CN107988385B (en) Method for detecting marker of PLAG1 gene Indel of beef cattle and special kit thereof
CN108060212B (en) DNA typing identification kit
CN102286476B (en) Short tandem repeat (STR) parting standard substance
CN110241234B (en) Fluorescence-labeled 32-plex InDels composite amplification system and application thereof
CN105316320B (en) DNA label, PCR primer and application thereof
CN108060228A (en) A kind of detection primer, kit and method for detecting BRCA1 and BRCA2 genetic mutations
CN105296471B (en) DNA label, PCR primer and application thereof
CN114657265B (en) SNP marker for identifying weaning weight of alpine merino sheep and application thereof
CN102286477B (en) Method for preparing short tandem repeat (STR) parting standard substances
CN109762909A (en) A kind of 44 site InDels composite amplification detection kits for sample medical jurisprudence individual appreciation of degrading
CN114657261A (en) SNP molecular marker related to thoracic vertebra number of sheep, primer group, kit, detection method and application
CN113373241A (en) Microsatellite marker of fishes in loach, and amplification primer and application thereof
CN108531609B (en) Set of golden monkshood EST-SSR primers and kit developed based on transcriptome sequencing
CN112266952A (en) STR (short tandem repeat) site supplementing composite amplification kit for difficult and complicated material to be detected and application of kit
CN108060233B (en) Fluorescence multiplex amplification system and kit for 30Y chromosome STR loci and application
Chandra et al. Genetic polymorphism of 21 STR markers in the representative sample of Indian population

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant
TR01 Transfer of patent right
TR01 Transfer of patent right

Effective date of registration: 20210107

Address after: 510700 4th floor, No.2 reagent workshop, 71 Fenghuang SANHENG Road, Zhongxin knowledge city, Jiulong Town, Huangpu District, Guangzhou City, Guangdong Province

Patentee after: GUANGZHOU KAIPU PHARMACEUTICAL TECHNOLOGY Co.,Ltd.

Patentee after: Shanghai Kaipu medical laboratory Co.,Ltd.

Address before: No.71, SANHENG Road, Fenghuang, Zhongxin knowledge city, Huangpu District, Guangzhou, Guangdong 510700

Patentee before: GUANGZHOU KAIPU PHARMACEUTICAL TECHNOLOGY Co.,Ltd.

TR01 Transfer of patent right
TR01 Transfer of patent right

Effective date of registration: 20210413

Address after: 510700 4th floor, No.2 reagent workshop, 71 Fenghuang SANHENG Road, Zhongxin knowledge city, Jiulong Town, Huangpu District, Guangzhou City, Guangdong Province

Patentee after: GUANGZHOU KAIPU PHARMACEUTICAL TECHNOLOGY Co.,Ltd.

Patentee after: Shanghai Kaipu medical laboratory Co.,Ltd.

Patentee after: HYBRIBIO Ltd.

Address before: 510700 4th floor, No.2 reagent workshop, 71 Fenghuang SANHENG Road, Zhongxin knowledge city, Jiulong Town, Huangpu District, Guangzhou City, Guangdong Province

Patentee before: GUANGZHOU KAIPU PHARMACEUTICAL TECHNOLOGY Co.,Ltd.

Patentee before: Shanghai Kaipu medical laboratory Co.,Ltd.