CN106480211A - A kind of kit and its SNP mark for detection of testis cancer neurological susceptibility - Google Patents
A kind of kit and its SNP mark for detection of testis cancer neurological susceptibility Download PDFInfo
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Abstract
The invention discloses a kind of SNP mark for detection of testis cancer neurological susceptibility, 16 SNP site are rs2900333, rs210138, rs7040024, rs755383, rs17021463, rs995030, rs3782181, rs4474514, rs12699477, rs4624820, rs7221274, rs4888262, rs4635969, rs9905704, rs3790665 and rs4657482.Pcr amplification primer thing and Single base extension primer and its kit of SNP mark is also disclosed, is that risk assessment, the diagnosis reference of carcinoma of testis provides important evidence.
Description
Technical field
The invention belongs to Genetic Detection field, more particularly to a kind of kit for detection of testis cancer neurological susceptibility and its
SNP mark.
Background technology
Using SNP (Single Nucleotide Polymorphsm, SNP) as genomic marker
Association analysis method is one of inheritance susceptible gene tester of disease the most frequently used at present.SNP is referred in genomic level
The DNA sequence polymorphism caused by single nucleotide variations, the occurrence frequency in crowd are more than 1%, and it is that the mankind are heritable
Modal one kind in variation.SNP genetic stability is strong, it is easy to detect, the SNP positioned at gene internal can directly influence albumen
The structure of matter or expression, and then have influence on tissue, organ or even physiological function.
Early detection is carried out to the related multiple genetic markers of common disease and system evaluation will be helpful to the early stage to disease
Prevention, diagnosis and treatment.At present, research has been found that science of heredity mark related to various common diseases in a large number, however, due to
Lacking has enough sensitivity and can multiple disease correlated identities be carried out with extensively (high flux detection site, high flux detection
Sample) method of the examination with checking so that the science of heredity mark of these common diseases extensively cannot be applied.Additionally, common at present
Disease genetic mark shortage system, effectively integrate, constrain significantly sending out in terms of common disease early prevention, diagnosis and treatment
Exhibition.Existing detection only has the genetic marker detection of single kind of disease or a class disease, and detection range is limited, and some common diseases
The method that there is no early detection.
At present, the detection of some traditional medicine means, such as histocyte has its intrinsic defect, position of drawing materials is improper,
Histocyte sample material is not enough or thinks to lack experience etc. will all cause carcinoma of testis mistaken diagnosis.Although other technologies such as iconography
Be widely used in inspection and the diagnosis of carcinoma of testis, but its disease carcinoma of testis degree qualitative on there are still very big office
Sex-limited.
In sum, a kind of kit for by multiple susceptibility loci detection of testis cancer neurological susceptibilities is researched and developed to carcinoma of testis
The prediction of onset risk, prevention, diagnosis are provided according to significant.
Content of the invention
In view of the defect that above-mentioned prior art is present, the purpose of the present invention is to propose to a kind of be used for detection of testis cancer neurological susceptibility
SNP mark, the pcr amplification primer thing of SNP mark and Single base extension primer and its kit present invention 16 SNP positions
Point is risk assessment, the diagnosis reference offer important evidence of carcinoma of testis.
The purpose of the present invention will be achieved by the following technical programs：
A kind of SNP mark for detection of testis cancer neurological susceptibility, the SNP mark include 16 SNP site, described
16 SNP site are rs2900333, rs210138, rs7040024, rs755383, rs17021463, rs995030,
rs3782181、rs4474514、rs12699477、rs4624820、rs7221274、rs4888262、rs4635969、
Rs9905704, rs3790665 and rs4657482.
The pcr amplification primer thing of above-mentioned SNP mark and Single base extension primer,
The sequence of the pcr amplification primer thing of the detection rs2900333 is respectively SEQ ID NO：1 and SEQ ID NO：2, with
And the sequence of Single base extension primer is SEQ ID NO：3；
The sequence of the pcr amplification primer thing of the detection rs210138 is respectively SEQ ID NO：4 and SEQ ID NO：5, with
And the sequence of Single base extension primer is SEQ ID NO：6；
The sequence of the pcr amplification primer thing of the detection rs7040024 is respectively SEQ ID NO：7 and SEQ ID NO：8, with
And the sequence of Single base extension primer is SEQ ID NO：9；
The sequence of the pcr amplification primer thing of the detection rs755383 is respectively SEQ ID NO：10 and SEQ ID NO：11,
And the sequence of Single base extension primer is SEQ ID NO：12；
The sequence of the pcr amplification primer thing of the detection rs17021463 is respectively SEQ ID NO：13 and SEQ ID NO：
14, and the sequence of Single base extension primer be SEQ ID NO：15；
The sequence of the pcr amplification primer thing of the detection rs995030 is respectively SEQ ID NO：16 and SEQ ID NO：17,
And the sequence of Single base extension primer is SEQ ID NO：18；
The sequence of the pcr amplification primer thing of the detection rs3782181 is respectively SEQ ID NO：19 and SEQ ID NO：20,
And the sequence of Single base extension primer is SEQ ID NO：21；
The sequence of the pcr amplification primer thing of the detection rs4474514 is respectively SEQ ID NO：22 and SEQ ID NO：23,
And the sequence of Single base extension primer is SEQ ID NO：24；
The sequence of the pcr amplification primer thing of the detection rs12699477 is respectively SEQ ID NO：25 and SEQ ID NO：
26, and the sequence of Single base extension primer be SEQ ID NO：27；
The sequence of the pcr amplification primer thing of the detection rs4624820 is respectively SEQ ID NO：28 and SEQ ID NO：29,
And the sequence of Single base extension primer is SEQ ID NO：30；
The sequence of the pcr amplification primer thing of the detection rs7221274 is respectively SEQ ID NO：31 and SEQ ID NO：32,
And the sequence of Single base extension primer is SEQ ID NO：33；
The sequence of the pcr amplification primer thing of the detection rs4888262 is respectively SEQ ID NO：34 and SEQ ID NO：35,
And the sequence of Single base extension primer is SEQ ID NO：36；
The sequence of the pcr amplification primer thing of the detection rs4635969 is respectively SEQ ID NO：37 and SEQ ID NO：38,
And the sequence of Single base extension primer is SEQ ID NO：39；
The sequence of the pcr amplification primer thing of the detection rs9905704 is respectively SEQ ID NO：40 and SEQ ID NO：41,
And the sequence of Single base extension primer is SEQ ID NO：42；
The sequence of the pcr amplification primer thing of the detection rs3790665 is respectively SEQ ID NO：43 and SEQ ID NO：44,
And the sequence of Single base extension primer is SEQ ID NO：45；
The sequence of the pcr amplification primer thing of the detection rs4657482 is respectively SEQ ID NO：46 and SEQ ID NO：47,
And the sequence of Single base extension primer is SEQ ID NO：48.
A kind of kit for detection of testis cancer neurological susceptibility, the kit include SNP mark described in claim 2
Pcr amplification primer thing and Single base extension primer, for detect in peripheral blood DNA 16 SNP site be rs2900333,
rs210138、rs7040024、rs755383、rs17021463、rs995030、rs3782181、rs4474514、
Rs12699477, rs4624820, rs7221274, rs4888262, rs4635969, rs9905704, rs3790665 and
rs4657482.
A kind of kit for detection of testis cancer neurological susceptibility, the kit also include Taq enzyme, dNTP mixed liquor,
MgCl2Solution, PCR reaction buffer, endonuclease reaction buffer solution, deionized water.
Kit of the present invention is the carcinoma of testis crowd with the present inventor based on multiple international and national large sample amounts
Based on the early-stage Study of normal population testis cancer susceptibility gene screening results, by the carcinoma of testis of the present inventor's autonomous Design
SNP screening process, screens qualified SNP site from NCBI-pubmed database：1) text related to carcinoma of testis is searched
Offer, by factor of influence and deliver the time limit and filtered；2) summary of candidate's document is checked, is excluded and carcinoma of testis SNP further
The unrelated document of research；3) document is read, finds the corresponding SNP site of carcinoma of testis；4) with the SNP site of selection and carcinoma of testis it is
Keyword consulting literatures again；5) judge whether the SNP site that chooses is closely related with carcinoma of testis, choose most close with carcinoma of testis
SNP site, corresponding OR value and mutating alkali yl that cut is closed；6) SNP site that previous step is obtained, public by Sequenom
Taking charge of software Typer 4.0 carries out online evaluation, obtains 16 final SNP site lists.
Compared with prior art, the invention provides a kind of detection method and reagent for detection of testis cancer neurological susceptibility
Box, has also reached following effect：16 SNP site of the present invention can be used for testis through mcta analysis with higher practicality
The earlier evaluations of ball cancer and extensive examination, and this 16 sites detection success rate is high, technique reproducible is good, cost performance is high,
For the risk assessment of carcinoma of testis, diagnosis with reference to important evidence is provided, to realize the earlier evaluations to carcinoma of testis disease.
Below just in conjunction with the embodiments, the specific embodiment to the present invention is described in further detail, so that technical scheme is more
Should be readily appreciated that, grasp.
Specific embodiment
Below by specific embodiment, the present invention will be described, but the invention is not limited in this.In following embodiments
The experimental technique, if no special instructions, is conventional method；The reagent and material, if no special instructions, all can be from business
Approach is obtained, and example below is simultaneously not used to limit the scope of the claims of the present invention, all equivalence enforcements without departing from carried out by the present invention
Or change, it is intended to be limited solely by this patent protection domain.
Embodiment 1 is used for the related SNP site of detection of testis cancer neurological susceptibility
Wherein, rs2900333 is located at Gene A TF7IP region, and rs210138 is located at gene BAK1 region, is rs7040024
With rs755383 be located at gene DMRT1 region, rs17021463 be located at gene HP GDS region, rs995030, rs3782181 and
Rs4474514 is located at gene KITLG region, and rs12699477 is located at gene M AD1L1 region, and rs4624820 is located at gene
NDFIP1 region, rs7221274 are located at gene PPM1E region, and rs4888262 is located at gene RFWD3 region, rs4635969 position
In gene TERT region, rs9905704 is located at gene TEX14 region, and rs3790665 and rs4657482 is located at gene UCK2 area
Domain.
Embodiment 2 is used for the kit of detection of testis cancer neurological susceptibility
First, the preparation of kit：
1st, design and synthesize pcr amplification primer thing and the Single base extension primer of 16 SNP site.SNP site to be measured
Pcr amplification primer thing and Single base extension primer refer to table 1
The pcr amplification primer thing of the SNP site to be measured of table 1 and Single base extension primer
2nd, kit also includes Taq enzyme, dNTP mixed liquor, MgCl2Solution, PCR reaction buffer, endonuclease reaction buffering
Liquid, deionized water.
2nd, the detection method of kit：
1st, the extraction of DNA
1.1 buccal swab DNA are extracted
1) buccal swab is placed in 2ml centrifuge tube, adds 400 μ l PBS.
2) 20 μ l QIAGEN Protease and 400 μ lBuffer AL are added.Vortex 15s is mixed immediately.In order to ensure have
The cracking of effect, sample and Buffer AL immediately and must be sufficiently mixed.
3) 56 DEG C of incubation 10min.Of short duration centrifugation.
4) add 400 μ l ethanol (96-100%) in sample, be vortexed and mix.Of short duration centrifugation is so that no liquid in lid.
5) liquid is transferred to centrifugal column, 8000rpm (6000 × g) is centrifuged 1min.
6) pillar is put into new 2ml EP pipe, plus 500 μ l Buffer AW1,8000rpm centrifugation 1min, abandons EP pipe and gives up
Liquid.
7) pillar is put into new 2mlEP pipe, plus 500 μ l Buffer AW2,14000rpm (20,000 × g) centrifugation 3min,
Abandon EP pipe and waste liquid.
8) pillar is put into new 2mlEP pipe, 14000rpm blank pipe is centrifuged 1min, abandons EP pipe.
9) new 1.5mlEP pipe, plus 120 μ l Buffer AE are put into pillar, incubated at room 5min, 8000rpm are centrifuged
1min, -20 DEG C of preservations.
1.2 whole blood DNAs are extracted
1) 20 μ l QIAGEN Protease are added in 1.5ml EP pipe.
2) to 200 μ l whole blood sample of Guan Zhongjia.Note：The purpose for being initially charged enzyme is to ensure the mixing of enzyme and blood.
3) add 200 μ l Buffer AL, be vortexed and mix 15s.
4) 56 DEG C of incubation 10min, of short duration centrifugation.Note：Extending incubation time can not increase yield or improve quality.
5) 200 μ l absolute ethyl alcohols are added, after vortex 15s, of short duration centrifugation is so that no liquid in lid.
6) continue 5～10 step of buccal swab.
2nd, PCR amplification
2.1 prepare pcr amplification reaction system in a new 1.5mlEP pipe, are shown in Table 2
2 pcr amplification reaction system of table
Above reagent adds 2 μ l per hole after mixing.
2.2 sequentially add DNA 10ng/ul1 μ l, 0.25uM Primer Mix 2 μ l, 5 μ l of cumulative volume per hole.
2.3 set in the PCR instrument of compatible 96 orifice plate PCR reaction condition as：94 DEG C of 4min, 94 DEG C of 20s, 56 DEG C
30min, 72 DEG C of 1min, 45 circulations；72℃ 3min；4 DEG C of holdings.96 hole PCR reaction plates are positioned in PCR instrument, are started
PCR reacts.
3rd, PCR primer alkaline phosphatase treatment
3.1 by PCR primer SAP (shrimp alkaline phosphatase, shrimp alkalescence phosphorus after PCR reaction terminates
Sour enzyme) process, with remove system middle reaches from dNTPs.
3.2 prepare alkaline phosphatase treatment reaction system, are shown in Table 3
3 alkaline phosphatase treatment reaction system of table
Above reagent adds 2 μ l per hole after mixing.
3.3 in the PCR instrument of compatible 96 orifice plate, sets PCR reaction condition：37℃ 40min；85℃ 5min；4 DEG C of guarantors
Hold, starting PCR instrument carries out alkaline phosphatase treatment.
4 Single base extensions
4.1 after alkaline phosphatase treatment terminates, and carries out single base extension.
4.2 prepare single base extension system, are shown in Table 4
4 single base extension system of table
Above reagent adds 1.06 μ l per hole after mixing
4.3 add iPLEX Extend Primer Mix 0.94 μ l, 9 μ l of cumulative volume per hole.
4.4 in the PCR instrument of compatible 96 orifice plate, sets PCR reaction condition：94 DEG C of 30s, 94 DEG C of 5s, 52 DEG C of 5s, 80
℃ 5s；4 circulations of 52 DEG C of 5s, 80 DEG C of 5s；94 DEG C of 5s, 52 DEG C of 5s, 80 DEG C of 5s, 39 circulations；72℃ 3min；4℃
Maintain.Starting PCR instrument carries out single base extension.
5th, purifying resin
5.1 tile Clean Resin resin in the resin plate of 6mg；
5.2 add 16 μ l water to the corresponding in the hole of extension products；
5.3 pour dried resin in extension products plate into, sealer, and slow speed vertical rotates 15min, makes resin and reaction
Thing is fully contacted；
5.4 3000rpm 5min centrifugation makes resin sink to bottom hole portion.
6th, chip point sample
Start 1000 point sample instrument of MassARRAY NanodispenserRS, the extension products after purifying resin are moved to 96
On the solid support of hole, chip is produced.
7th, Mass Spectrometer Method
Chip is used MALDI-TOF (matrix-assistedlaser desorption/ionization-time
Offlight), matrix solid-dispersion flight time mass spectrum) analysis, testing result is using TYPER4.0 software
(Sequenom) parting output result, analyze experimenter's carcinoma of testis occurrence risk, to realize the earlier evaluations to disease.
The document that embodiment 3 is quoted with 16 closely related SNP site of carcinoma of testis, is shown in Table 5
The document that table 5 is quoted with 16 closely related SNP site of carcinoma of testis
Embodiment 4 draws associating for 16 SNP site and cervical carcinoma according to the document that quotes in embodiment 3, is shown in Table 6
6 16 SNP site of table and the association analysis result of carcinoma of testis
The checking of 5 16 SNP site of embodiment
16 SNP site are analyzed using the Mass Spectrometry detection method of kit in above-described embodiment 2, testing result is used
16 SNP site of table 6 and testis in TYPER4.0 software (Sequenom) parting output result, its OR value and embodiment 4
The association analysis result of cancer is identical, illustrate 16 sites have higher practicality, can be used for carcinoma of testis earlier evaluations and
Extensive examination, as analytical technique of mass spectrum has, detection success rate is high, technique reproducible is good, cost performance is high, therefore the present invention
Use Mass Spectrometer Method to analyze this 16 sites detection success rate is high, technique reproducible is good, cost performance is high, is the trouble of carcinoma of testis
Sick risk assessment, diagnosis reference provide important evidence, to realize the earlier evaluations to carcinoma of testis disease.
Described above illustrate and describes some preferred embodiments of the present invention, but as previously mentioned, it should be understood that the present invention
Be not limited to form disclosed herein, be not to be taken as the exclusion to other embodiment, and can be used for various other combinations,
Modification and environment, and can be in invention contemplated scope described herein, by the technology or knowledge of above-mentioned teaching or association area
It is modified.And change that those skilled in the art are carried out and change be without departing from the spirit and scope of the present invention, then all should be at this
In the protection domain of bright claims.
SEQUENCE LISTING
<110>Nucleon Gene Tech. Company Limited of Shenzhen
<120>A kind of kit and its SNP mark for detection of testis cancer neurological susceptibility
<130>
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<170> PatentIn version 3.5
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Claims (4)
1. a kind of SNP mark for detection of testis cancer neurological susceptibility, it is characterised in that the SNP mark includes 16 SNP
Site, 16 SNP site be rs2900333, rs210138, rs7040024, rs755383, rs17021463,
rs995030、rs3782181、rs4474514、rs12699477、rs4624820、rs7221274、rs4888262、
Rs4635969, rs9905704, rs3790665 and rs4657482.
2. the pcr amplification primer thing of SNP mark according to claim 1 and Single base extension primer, it is characterised in that
The sequence of the pcr amplification primer thing of the detection rs2900333 is respectively SEQ ID NO:1 and SEQ ID NO:2, Yi Jidan
The sequence of base extension primer is SEQ ID NO:3；
The sequence of the pcr amplification primer thing of the detection rs210138 is respectively SEQ ID NO:4 and SEQ ID NO:5, Yi Jidan
The sequence of base extension primer is SEQ ID NO:6；
The sequence of the pcr amplification primer thing of the detection rs7040024 is respectively SEQ ID NO:7 and SEQ ID NO:8, Yi Jidan
The sequence of base extension primer is SEQ ID NO:9；
The sequence of the pcr amplification primer thing of the detection rs755383 is respectively SEQ ID NO:10 and SEQ ID NO:11, and
The sequence of Single base extension primer is SEQ ID NO:12；
The sequence of the pcr amplification primer thing of the detection rs17021463 is respectively SEQ ID NO:13 and SEQ ID NO:14, with
And the sequence of Single base extension primer is SEQ ID NO:15；
The sequence of the pcr amplification primer thing of the detection rs995030 is respectively SEQ ID NO:16 and SEQ ID NO:17, and
The sequence of Single base extension primer is SEQ ID NO:18；
The sequence of the pcr amplification primer thing of the detection rs3782181 is respectively SEQ ID NO:19 and SEQ ID NO:20, and
The sequence of Single base extension primer is SEQ ID NO:21；
The sequence of the pcr amplification primer thing of the detection rs4474514 is respectively SEQ ID NO:22 and SEQ ID NO:23, and
The sequence of Single base extension primer is SEQ ID NO:24；
The sequence of the pcr amplification primer thing of the detection rs12699477 is respectively SEQ ID NO:25 and SEQ ID NO:26, with
And the sequence of Single base extension primer is SEQ ID NO:27；
The sequence of the pcr amplification primer thing of the detection rs4624820 is respectively SEQ ID NO:28 and SEQ ID NO:29, and
The sequence of Single base extension primer is SEQ ID NO:30；
The sequence of the pcr amplification primer thing of the detection rs7221274 is respectively SEQ ID NO:31 and SEQ ID NO:32, and
The sequence of Single base extension primer is SEQ ID NO:33；
The sequence of the pcr amplification primer thing of the detection rs4888262 is respectively SEQ ID NO:34 and SEQ ID NO:35, and
The sequence of Single base extension primer is SEQ ID NO:36；
The sequence of the pcr amplification primer thing of the detection rs4635969 is respectively SEQ ID NO:37 and SEQ ID NO:38, and
The sequence of Single base extension primer is SEQ ID NO:39；
The sequence of the pcr amplification primer thing of the detection rs9905704 is respectively SEQ ID NO:40 and SEQ ID NO:41, and
The sequence of Single base extension primer is SEQ ID NO:42；
The sequence of the pcr amplification primer thing of the detection rs3790665 is respectively SEQ ID NO:43 and SEQ ID NO:44, and
The sequence of Single base extension primer is SEQ ID NO:45；
The sequence of the pcr amplification primer thing of the detection rs4657482 is respectively SEQ ID NO:46 and SEQ ID NO:47, and
The sequence of Single base extension primer is SEQ ID NO:48.
3. a kind of kit for detection of testis cancer neurological susceptibility, it is characterised in that the kit is included described in claim 2
The pcr amplification primer thing of SNP mark and Single base extension primer, for detecting that 16 SNP site are in peripheral blood DNA
rs2900333、rs210138、rs7040024、rs755383、rs17021463、rs995030、rs3782181、
rs4474514、rs12699477、rs4624820、rs7221274、rs4888262、rs4635969、rs9905704、
Rs3790665 and rs4657482.
4. a kind of kit for detection of testis cancer neurological susceptibility that is stated according to claim 3, it is characterised in that the kit
Also include Taq enzyme, dNTP mixed liquor, MgCl2Solution, PCR reaction buffer, endonuclease reaction buffer solution, deionized water.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201611049653.4A CN106480211A (en) | 2016-11-24 | 2016-11-24 | A kind of kit and its SNP mark for detection of testis cancer neurological susceptibility |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201611049653.4A CN106480211A (en) | 2016-11-24 | 2016-11-24 | A kind of kit and its SNP mark for detection of testis cancer neurological susceptibility |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110241227A (en) * | 2019-07-01 | 2019-09-17 | 兰州大学 | A kind of method and application detecting sheep SPATA6 gene single nucleotide polymorphism |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101641451A (en) * | 2006-10-27 | 2010-02-03 | 解码遗传学私营有限责任公司 | Cancer susceptibility variants on the chr8q24.21 |
| CN102137937A (en) * | 2008-07-09 | 2011-07-27 | 解码遗传学私营有限责任公司 | Genetic variants as markers for use in urinary bladder cancer risk assessment, diagnosis, prognosis and treatment |
-
2016
- 2016-11-24 CN CN201611049653.4A patent/CN106480211A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101641451A (en) * | 2006-10-27 | 2010-02-03 | 解码遗传学私营有限责任公司 | Cancer susceptibility variants on the chr8q24.21 |
| CN102137937A (en) * | 2008-07-09 | 2011-07-27 | 解码遗传学私营有限责任公司 | Genetic variants as markers for use in urinary bladder cancer risk assessment, diagnosis, prognosis and treatment |
Non-Patent Citations (1)
| Title |
|---|
| KEVIN LITCHFIELD等: ""The genomic landscape of testicular germ cell tumours: from susceptibility to treatment"", 《NATURE REVIEWS》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110241227A (en) * | 2019-07-01 | 2019-09-17 | 兰州大学 | A kind of method and application detecting sheep SPATA6 gene single nucleotide polymorphism |
| CN110241227B (en) * | 2019-07-01 | 2022-05-17 | 兰州大学 | Method for detecting sheep SPATA6 gene single nucleotide polymorphism and application |
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