CN103820564A - Multiplex amplification kit for 25 STRs (short tandem repeats) - Google Patents
Multiplex amplification kit for 25 STRs (short tandem repeats) Download PDFInfo
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- CN103820564A CN103820564A CN201410091492.XA CN201410091492A CN103820564A CN 103820564 A CN103820564 A CN 103820564A CN 201410091492 A CN201410091492 A CN 201410091492A CN 103820564 A CN103820564 A CN 103820564A
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- 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
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
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- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
Abstract
The invention relates to a multiplex amplification system for analyzing a plurality of STR (short tandem repeat) gene loci simultaneously. The system is characterized in that 25 gene loci are subjected to multiplex amplification and include: 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 further relates to a method and a kit for analyzing DNA (deoxyribonucleic acid) samples and applications of the method and the kit.
Description
Technical field:
The present invention relates to biological technical field.Further, the present invention relates to detect the genetic marker gene in human genome with polymorphism.The present invention be more particularly directed to multiple short tandem repeats that simultaneously increase with polymerase chain reaction in an amplification system.
Background technology:
STR (being called for short STR) repeats (SSR) also referred to as micro-satellite or simple sequence, is that a class is extensively present in the strand of dna connection tumor-necrosis factor glycoproteins in eukaryotic gene group, and core sequence is 2-6 base repeating unit.Str locus seat quantity is large, branch is extensive, account for whole genomic 3% (Intemational Human Genome Sequencing Consortium, 2001), and polymorphism is high, its polymorphism is mainly derived from the difference of core sequence multiplicity between individuality, and this species diversity is followed mendelian inheritance in genetic process.Therefore, STR augmentation detection technology is widely used in individual recognition, relationship is identified and population genetic study.
STR composite amplification technology is the technical way of legal medical expert's individual recognition and paternity test at present, and be used widely (" Forensic DNA Typing " second edition, John Butler) in the DNA laboratory all over the world.In case analysis, be used widely, for cracking of cases provides strong evidence.Along with development, a lot of countries utilize this technology to set up offender and suspect's DNA database, be namely in prison convict and suspect's DNA data analysis are also entered in database, are convenient to compare and the work such as investigation.
Early stage STR composite amplification technology can realize the str locus seat of 10 left and right of increasing in a reaction system, along with the increase of the extensive and comparing of applying, the information that 10 locus provide can not meet the demands, producer both domestic and external develops the product of the more polygene seat making new advances, such as the AmpFISTR Identifiler test kit of American AB I company and Promega company of the U.S.
16 test kits are all that 15 str locus seats add sex determining gene.Domestic also have a similar product, such as the DNATyper15 of two of the Ministry of Public Security can realize 15 locus amplifications (research of DNATyper15 locus and selection, Ye Jian etc., " criminal technique ", 2007 03) simultaneously.
But in recent years along with DNA is more and more wider as the application of identification of means, user has had higher requirement to locus quantity, information content and the proliferation time of test kit.Aspect DNA database establishment, for test kit compatibility, new requirement is also proposed.In the current DNA database of China, there are 2,000,000 parts of left and right, China will further accelerate aspect DNA database establishment, to the data in DNA database in 2013 by 10 million part, in the database of the developed countries such as Britain, approximately have account for total population 10% data (
http:// www.forensic.gov.uk.Asplen, 2004).Along with China DNA database establishment scale will constantly expand, the effect of comparing is more and more important.Comparing is to be based upon on the data basis that the analysis of STR composite amplification obtains, need each STR test kit on locus, to there is compatibility (forensic DNA database in Chian, " Chinese law medical journal ", Jiang Xianhua, the 12nd the 5th phase of volume in 2006).
13 core gene seats (vWA, D21S11, D18S51, D5S818, D7S820, D13S317, D16S539, FGA, D8S1179, D3S1358, CSF1PO, THO1, TPOX) that currently used test kit stipulates mainly with the CODIS standard of the U.S. are greatly basis, just select to increase other locus on this basis, also have portioned product to delete the portion gene seat in 13 core sequences.But along with the locus quantity of user to test kit, information content and compatible higher demand, each manufacturer has released the test kit that has multidigit point more and meet up-to-date CODIS site.Each manufacturer test kit locus information is in table 1.So if adopt different manufacturers' STR test kit, the data that import in DNA database have locus difference.In comparison, not all data are all effective like this, and just the data of homologous genes seat part can be compared, and distinct portions just cannot be applied.For example, Identifiler test kit and
between 16 test kits, identical str locus seat is 13, so other two str locus seats do not act on when the comparing due to different.And these 13 can be compared locus data is reliably for getting rid of when, but for inadequate often in situation about assert.Need more locus data.So, due to each STR test kit locus difference, caused the waste of DNA database section data resource, and validity is inadequate.
Along with STR detection kit is more and more extensive in the application of client unit, rapid amplifying is also an aspect of test kit performance boost, and amplification rate has greatly reduced the waste of time cost faster, has improved working efficiency.Traditional test kit proliferation time is about 3 to 4 hours, and the previous generation Goldeneye20A of our company test kit proliferation time is also about 2.5 hours.If can realize in 1 hour and complete amplified reaction one time, can greatly promote the performance of test kit, also can increase work efficiency.
Therefore, the DNA field of identifying need to have a reaction increase polygene seat more, more information amount can be provided, there is the STR composite amplification body of better compatibility and faster amplification rate.If can realize more than 25 locus of primary first-order equation amplification in one hour, not only can save reagent cost, can also increase work efficiency, save human cost etc.
Table 1: the locus information of each main flow manufacturer's test kit
Note: black font represents CODIS13 locus ,+represent the locus that is included ,-locus not to be covered represented
Summary of the invention:
The present invention be directed to the demand, model the composite amplification system of 25 str locus seats of one-time detection, these str locus seats have comprised the full gene seat that each manufacturer adopts at present 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.
Amplification system of the present invention comprises primer mixture, reaction buffer and warm start Taq archaeal dna polymerase etc.
First design respectively Auele Specific Primer for above-mentioned 25 locus at the flank of its tumor-necrosis factor glycoproteins.Design of primers adopts Primer5 software, and every primer annealing temperature is close to or higher than 60 ℃.Can not produce primer dimer, other interaction or cross reaction, amplified production length is between 90-480bp.Every pair of primer is carried out amplification assay and optimized, until obtain clear single amplified band.Primer sequence sees the following form 2.
The each locus primer sequence of table 2 composite amplification system
According to expanding fragment length etc., said gene seat is divided into 4 groups, first group comprises D22S1045, TPOX, D13S317, D7S820, D1S1656, Penta E and D10S1248, second group comprises D8S1179, D5S818, D19S433, D16S539, CSF1PO and Penta D, the 3rd group comprises D3S1358, vWA, D2S1338, D18S51, D6S1043 and D2S441, and the 4th group comprises Amelogenin, TH01, D12S391, D21S11, FGA and DYS391.Every group fluorescein-labelled by difference respectively, and each locus amplified production separates according to difference in length among every group, and two locus can not have overlapping.Respectively every group of primer carried out to composite amplification test.Determine that this group does not have non-specific amplification phenomenon, after the situations such as no cross reaction, adjust the concentration of every pair of primer, each fragment peak equalization in group is reached more than 40%.
4 groups of primers are used respectively to blueness, green, yellow and red fluorescence element mark.Chain of a mark in every pair of primer, is marked at 5 ' of primer and holds.Blue markings can be selected 5-FAM (CF), the fluorescein molecule of 6-FAM (6-Fluoresceincarboxylic acid) or close spectrum, Green Marker can be selected HEX (chlordene-6-methyl fluorescein), JOE (6-carboxyl-4,5-bis-chloro-2,7-dimethoxy fluorescein succinimide ester) or the fluorescein molecule of close spectrum, yellow mark can be selected the fluorescein molecule of TMR (4-methyl-6-carboxyl-rhodamine) or close spectrum, and red-label can be selected the fluorescein molecule of ROX (carboxyl-X-rhodamine) or close 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 reached more than 30%.The primer mixture obtaining can be for above-mentioned 25 locus composite amplifications.
Pcr amplification reaction of the present invention can carry out in certain buffer system.Buffer system comprises: 50mM KCI, 10mM Tris-HC1 (PH8.3,25 ℃), 2.0mM MgCl
2, the dNTP of 0.1mg/ml BSA (bovine serum albumin) and each 0.2mM.DNTP is the molar mixtures such as four kinds of deoxyribonucleotides (dATP, dTTP, dCTP, dGTP).
The Taq archaeal dna polymerase that reacts required is warm start archaeal dna polymerase, antibody sealing modify or chemically modified can.Each amplification system of the present invention (25 μ 1) needs the Taq archaeal dna polymerase of 2U to 4U.
Program below amplification system (as ABI9700, ABI9600, ABI2720, Bio-Rad iCycler, Bio-Rad C1000 etc.) on various reaction heat circulating instruments adopts can obtain good result: 96 ℃ are incubated 2 minutes; 98 ℃ were incubated for 2 seconds, and 60 ℃ were incubated for 40 seconds, and 68 ℃ were incubated for 20 seconds, 30 circulations of this step operation; 4-10 ℃ of insulation.
Template DNA in the present invention is human genome DNA.By various ordinary methods, the template DNA extracting such as methods (" molecular cloning laboratory manual " third edition, cold spring port press) such as paramagnetic particle method, phenol chloroform method, resin purifications all can obtain good result.DNA can be by preparing with undertissue or cell: blood (blood cake), seminal fluid (seminal stain), bone, hair, saliva (salivary stain), sweat, the amniotic fluid etc. that contains fetal cell.DNA profiling amount preferably can obtain good amplification in the scope of 0.25ng to 4ng, and template amount is too low may cause some locus inspection not measure, and the too high meeting of template amount causes nonspecific amplified production to produce.
In above-mentioned reaction buffer system, according to the response procedures amplification template DNA specifying, can obtain the amplified production that each locus mixes.The present invention is owing to having adopted fluorescently-labeled primer, amplified production is also with fluorescent marker, and marker can send the optical signal that can pass through sequenator (as ABI377,310DNA sequencer) or genetic analyzer (as ABI3130,3100genetic analyzer) identification under laser excitation, so amplified production can be by carrying out electrophoresis and detecting and analyze on the instrument such as sequenator or genetic analyzer.
In detecting on sequenator or genetic analyzer, in amplified production and molecular weight, mark (marker, intemal lane standard), methane amide mix according to a certain percentage, enter electrophoretic separation in instrument kapillary or gel.In molecular weight, mark is made up of the fluorescent label DNA fragment of many known length, is used for calculating pcr amplification product fragment length, thereby can judges gene type and compare with allelic ladder.
Data after electrophoresis can be analyzed in the data analysis software such as GeneMapper, GeneMarker, GeneScan, obtain str locus somatotype collection of illustrative plates and data.
The present invention relates to
(1) analyze the composite amplification systems of multiple str locus seats simultaneously, it is characterized in that: composite amplification 25 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.
(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 is D8S1179, D5S818, D19S433, D16S539, CSF1PO and Penta D; The 3rd group is D3S1358, vWA, D2S1338, D18S51, D6S1043 and D2S441; The 4th group is Amelogenin, TH01, D12S391, D21S11, FGA and DYS391, and wherein, first group of the primer of amplification is SEQ ID NO:1-14; Second group is SEQ ID NO:15-26; The 3rd group is SEQ ID NO:27-38; The 4th group is SEQ ID NO:39-50.
(3), according to the composite amplification system described in item 2,25 locus wherein increase in a composite amplification reaction system simultaneously.
(4) according to the composite amplification system described in item 2, wherein locus is positioned at the pair of primers amplification of these locus both sides, wherein in every pair of primer, has 5 ' end of a primer to carry out fluorescein-labelled.
(5), according to the composite amplification system described in item 1, wherein the multiple str locus seats of composite amplification utilize polymerase chain reaction to carry out.
(6) according to the composite amplification system described in item 2, wherein this system comprises primer mixture.
(7) method for while analyzing DNA sample, is characterized in that the composite amplification system of applying above-mentioned any one detects DNA.
(8), according to the method described in item 7, wherein DNA sample comprises in blood, blood cake, seminal fluid, seminal stain, bone, hair, saliva, salivary stain, sweat and amniotic fluid one or more.。
(9) shown in SEQ ID NO:1-50 for the primer sequence of composite amplification system or the mixture of described primer sequence.
(10) a kind of for analyze the test kit of multiple str locus seats 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, and this test kit comprises the primer shown in SEQ ID NO:1-50.
The present invention also provides to have and is selected from the primer sequence described in table 2, wherein one or more, preferably 1-15, preferably 1-10, preferably 1-5 the modified sequence that Nucleotide is substituted, deletes and/or add 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 are for analyzing the purposes of multiple str locus seats.
Adopt the present invention only to need one-time detection, operating time, detection time and detection reagent cost all reduce 50% or more.Adopt the present invention to there is very high compatibility, use the present invention not worry the problem of data compatibility before.
Accompanying drawing explanation
Fig. 1-1 and Fig. 1-2 are two portions that DNA sample adopts the analysis collection of illustrative plates after the present invention's amplification, and the right of Fig. 1-1 and the left side of Fig. 1-2 have overlapping, and both can amalgamation be a complete collection of illustrative plates.
Fig. 2-1 and Fig. 2-2 adopt the two portions of the analysis collection of illustrative plates after the cognitive Goldeneye20A of the company test kit amplification of basic point for DNA sample, and the right of Fig. 2-1 and the left side of Fig. 2-2 have overlapping, and both can amalgamation be a complete collection of illustrative plates.
Embodiment
Mode below by embodiment further illustrates the present invention.The following examples are only used to the object of explanation, and not limit the scope of the invention.
25 locus of embodiment 1 composite amplification are also analyzed its genotype
Blood is contributed by volunteer.Template DNA, by extracting by chelex-100 method in blood, separately adds legal medical expert's somatotype standard substance 9948.Amplified reaction carries out on ABI9700 thermal cycler, and electrophoresis and detection are carried out on ABI3100 genetic analyzer, and data analysis adopts GeneMapper ID v3.2 software.Sample adopts the SinoFiler test kit of ABI company and the Goldeneye20A test kit of the cognitive company of basic point to detect simultaneously, and working method is carried out according to test kit specification sheets, and result in contrast.The reagent that the present invention is used and material such as allelic ladder (ladder) is the conventional conventional material of those skilled in the art.
1.1.chelex-100 method is extracted DNA (concrete grammar is with reference to " Forensic DNA Protocol ", Humana Press, 1998)
1) get blood that 3 μ l add antithrombotics in 500 μ l centrifuge tubes
2) vibration mixes chelex solution, chelex is fully suspended, and every pipe adds the chelex-100 (100-200mesh, purchased from Bio-Rad company) of 195 μ l5%, add again 5 μ l Proteinase Ks (20mg/ml, purchased from Tian Gen biochemical technology company limited)
3) oscillation sample, on constant-temperature metal bath, 56 ℃ of temperature were bathed after 2 hours, took out sample vibration 2 minutes,
4) boil 8-10 minute, centrifugal 3 minutes of 13000rpm
5) careful sucking-off approximately 150 μ l supernatants, are transferred in new pipe, and 10 μ l PCR reaction systems are got 1 μ l as template
1.2. polymerase chain reaction (PCR) amplification
1) get damping fluid, primer mixture, Taq enzyme, be made into mixed solution according to following table, after vibration mixes, divide and be filled in PCR reaction tubes, every pipe 25 μ l, add template DNA
2) according to reaction conditions below, thermal cycler is set and increases instrument (ABI9700PCR instrument), PCR reaction tubes is put into instrument and start amplification gene fragment.96 ℃ are incubated 2 minutes; 98 ℃ were incubated for 2 seconds, and 60 ℃ were incubated for 40 seconds, and 68 ℃ were incubated for 20 seconds, move 30 circulations; 4 ℃ continue insulation, until take out sample
1.3. after amplified reaction finishes, take out reaction tubes, carry out electrophoresis and detection with ABI3100 genetic analyzer
1) get (mark+10 μ l deionized formamides in 0.5 μ l molecular weight) × (sample number) and be made into mixed solution
2) mix rear packing, every pipe 10 μ l, then add respectively 1 μ l amplified production and allelic ladder (ladder), brief centrifugal by liquid collecting to centrifuge tube pipe bottom
3) 95 ℃ of sex change of sample 4 minutes, then rapid cooled on ice 4 minutes, makes the complete sex change of DNA and keeps denatured state
4) sample is put into the sample tray of genetic analysis instrument, instrument parameter (sample introduction voltage 3kV, sample injection time 10 seconds) is set, start electrophoresis detection
5) after about 40 minutes, electrophoresis finishes, and obtains collection of illustrative plates and somatotype result (seeing Fig. 1-1, Fig. 1-2, Fig. 2-1, Fig. 2-2 and table 3) by GeneMapper software analysis experimental data
The increase gene type result of same sample of two kinds of test kits of table 3
Invention effect:
Along with the increase of complex locus number, due to the impact of competition, the relative equilibrium control difficulty of each locus strengthens, the present invention is by experiment repeatedly, model the composite amplification system of 25 str locus seats of one-time detection, these str locus seats have been integrated most locus that each manufacturer adopts at present both at home and abroad.Utilize this amplification system can disposable detection 25 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.
Utilize this system single job to obtain 25 locus information, therefore this system has very high individual recognition rate, be equivalent to the information summation of simultaneously utilizing 2-3 other like products to obtain, no matter at pcr amplification and genetic analyzer detection, all provide cost savings and manpower, improved working efficiency.In amplification link, by increasing and be reduced to the amplification of a kind of test kit with first two or 3 kinds of test kits, save more than 50% reagent cost, time shorten 50%.Detection, adopts 2 kinds or 3 kinds of test kits need to distinguish sample detection, adopts the present invention only to need one-time detection, and operating time, detection time and detection reagent cost all reduce 50% or more.
Adopt the present invention to there is very high compatibility, use the present invention not worry the problem of data compatibility before.Because these 25 locus have comprised the current full gene seat of main product domestic and that use before, there is good compatibility, existing all data in not only can compatible China DNA database at present, and new generation product is also had to very high compatibility.
Be understandable that, description is above only example of the present invention, thereby the scope that the claims in the present invention are protected not merely limits with particular disclosed herein.Any embodiment being equal to will be deemed to be within the scope of the present invention.In fact, according to description above, it will be all possible for those skilled in the art that the present invention is carried out to relevant modifications and variations, thereby this modifications and variations are also by within dropping on the scope of the appended claim of the present invention.
Claims (10)
1. a composite amplification system of simultaneously analyzing multiple str locus seats, is characterized in that: composite amplification 25 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.
2. composite amplification system according to claim 1, wherein said locus is divided into following combination: first group is D13S317, TPOX, D22S1045, D7S820, D1S1656, Penta E and D10S1248; Second group is D8S1179, D5S818, D19S433, D16S539, CSF1PO and Penta D; The 3rd group is D3S1358, vWA, D2S1338, D18S51, D6S1043 and D2S441; The 4th group is Amelogenin, TH01, D12S391, D21S11, FGA and DYS391, and wherein, first group of the primer of amplification is SEQ ID NO:1-14; Second group is SEQ ID NO:15-26; The 3rd group is SEQ ID NO:27-38; The 4th group is SEQ ID NO:39-50.
3. composite amplification system according to claim 2,25 locus wherein increase in a composite amplification reaction system simultaneously.
4. composite amplification system according to claim 2, wherein locus is positioned at the pair of primers amplification of these locus both sides, wherein in every pair of primer, has 5 ' end of a primer to carry out fluorescein-labelled.
5. composite amplification system according to claim 1, wherein the multiple str locus seats of composite amplification utilize polymerase chain reaction to carry out.
6. composite amplification system according to claim 2, wherein this system comprises primer mixture.
7. a method for while analyzing DNA sample, is characterized in that application rights requires the described composite amplification system of one of 1-6 to detect DNA.
8. method according to claim 7, wherein DNA sample comprises one or more in blood, blood cake, seminal fluid, seminal stain, bone, hair, saliva, salivary stain, sweat and amniotic fluid.
Shown in 9.SEQ ID NO:1-50 for the primer sequence of composite amplification system or the mixture of described primer sequence.
10. one kind for analyze the test kit of multiple str locus seats 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, and this test kit comprises the primer shown in SEQ ID NO:1-50.
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| CN106011229A (en) * | 2016-04-26 | 2016-10-12 | 深圳华大法医科技有限公司 | Composite amplification system for human 18 STR sites, kit and use of composite amplification system and kit |
| 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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| MASAKI HASHIAYDA ET AL.: "Genetic variation in a Japanese population, using the multiplex 24 STRs analysis system", 《FORENSIC SCIENCE INTERNATIONAL: GENETICS SUPPLEMENT SERIES》 * |
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| CN105368958A (en) * | 2015-12-15 | 2016-03-02 | 基点认知技术(北京)有限公司 | Kit for realizing multiple amplification of 21 short tandem repeats by adopting degenerate primers |
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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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