CN109207625A - A kind of SSR-PCR identification method for distinguishing 4 main breeds of agaricus bisporus - Google Patents
A kind of SSR-PCR identification method for distinguishing 4 main breeds of agaricus bisporus Download PDFInfo
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Abstract
The invention belongs to edible mushroom molecular mark fields, and in particular to a kind of SSR-PCR identification method for distinguishing 4 main breeds of agaricus bisporus.This method carries out PCR specific amplified to agaricus bisporus country main breed As2796, W192, good fortune mushroom 38 and A15 using SSR molecular marker, generates specific manner.Method of the invention does not have to lengthy and jumbled economical character fruiting comparative test, the AFLP system of specificity can identify the main breed for distinguishing the agaricus bisporus country, this method time-consuming is short, it is easy to operate, high specificity, this method can rapidly identify 4 domestic agaricus bisporus main breeds in the production of agaricus bisporus strain, avoid the generation of homonymus or synonymum and xenogenesis phenomenon of the same name, improve the production efficiency of agaricus bisporus.
Description
Technical field
The invention belongs to edible mushroom molecular mark field, a kind of 4 main breeds of differentiation agaricus bisporus
The SSR-PCR identification method of As2796, W192, good fortune mushroom 38 and A15.
Background technique
Simple sequence repeats (simple sequence repeat, SSR), also known as microsatellite DNA, are with 2-6 alkali
Base is attached most importance to the tandem repetitive sequence of complex radical member, is widely present in the genome of higher organism.In numerous molecule labelling methods
In, the SSR marker based on genome has the characteristics that numerous specific site, codominance, answers equipotential, is widely distributed, has become at present
For building Genetic linkage map, research Population Genetics, draw Variety fingerprinting, the individual specific spectruming belt of screening and molecule mark
Remember the ideal tools of assistant breeding etc..
Agaricus bisporus (Agaricus bisporus) it is to cultivate most commonly used edible mushroom in the world, have extremely important
Economic value.In China, agaricus bisporus was also planted extensively, according to official statistics whole nation agaricus bisporus annual output 3380000 in 2016
Ton, accounts for 60% of Gross World Product or more, has become the emerging supporting industry in agricultural at present.Due to the production of China each department
Power disparate development, the production method for planting agaricus bisporus are also to show diversification, there is a full set of full machine for introducing the Netherlands pattern
The industrialized cultivation mode of tool also has the domestic mechanization production mode for being suitble to China domestic national conditions by improvement, also
The large number of small production workshop based on self-employed farmer.Currently, circulating in domestic agaricus bisporus main breed has
As2796, W192, good fortune mushroom 38 and A15 are investigated according to national mushroom industry technical system agaricus bisporus breed improvement post and are tied
Fruit, above-mentioned 4 main breed use ratios account for 90% or more.Wherein As2796 mushroom growing up time is more, and resistance to low level management is relatively suitble to small
Produce workshop plantation；W192 and 38 yield of good fortune mushroom are high, and quality is preferable, and fruiting is relatively concentrated, and are suitble to the factories in China kind of Improvement type
It plants；A15 yield is high, medium quality, and fruiting is concentrated, and is suitble to the Fully-mechanized industrialized cultivation mode of the Netherlands pattern.Due to not
Same planting patterns requires the extensive property of Different Varieties and cultivation management adaptable therewith also different, so producing in agaricus bisporus
Requirement in the process to strain authenticity is very high.But since agaricus bisporus can be with vegetative propagation, part peasant household and enterprise's kind
It plants family to separate frequently by tissue, expands numerous and conservation lack of standardizationly, during causing production and operation and managing circulation often
There is the phenomenon that homonymus or synonymum and xenogenesis of the same name, so being highly desirable to establish a set of quick identification to domestic main breed
Method.
My the assumed responsibility for sub- project of national science and technology supporting plan project " agaricus bisporus new varieties during 2013 to 2016
Cultivate and Key Techniques in Seed Production studied ", heredity is carried out to cultivation of agaricus bisporus bacterial strain both domestic and external by SSR molecular marker technology
Polymorphism analysis simultaneously establishes genetic relationship map.In the recent period, we explore on the Research foundation of the project through many experiments,
Exploitation obtains the SSR-PCR identification method of one group of identification agaricus bisporus As2796, W192, good fortune mushroom 38 and A15, utilizes the molecule
Label can rapidly be identified in the mycelia stage of bacterial strain, time saving and energy saving without being determined by fruiting according to economical character,
This has great significance to the authenticity identification of strain or kind in agaricus bisporus production process.
Summary of the invention
The present invention is based on molecular marking techniques, provide a kind of identification side SSR-PCR for distinguishing 4 main breeds of agaricus bisporus
Method.
A kind of SSR-PCR identification method of 4 main breeds of differentiation agaricus bisporus of the invention is to utilize SSR molecule mark
Note carries out PCR amplification to domestic agaricus bisporus main breed As2796, W192, good fortune mushroom 38 and A15.
SSR-PCR, different kind energy are carried out using the genomic DNA of primer pair country agaricus bisporus main breed
Amplify the map banding pattern product of specificity, the core former times acid sequence of the special primer are as follows:
Scaf4_1k11F:5 '-CAATCACAATCCCTCCAAGC-3 ',
Scaf4_1k11R:5 '-AAACGAATTAGGAACGGTGG-3 '；
Using above-mentioned primer pair Scaf4_1k11F and Scaf4_1k11R to agaricus bisporus As2796, W192, good fortune mushroom 38
SSR-PCR amplification is carried out with the genomic DNA of A15 bacterial strain, specifically AFLP system is that strains A s2796 can be amplified
The product of 1004bp；Bacterial strain W192 can amplify the product of 801bp；Bacterial strain good fortune mushroom 38 can amplify the production of 1004bp and 801bp
Object；Strains A 15 can amplify the product of 1004bp, 801bp and 252bp.
The SSR-PCR amplification condition are as follows: 94 DEG C of initial denaturation 5min；94 DEG C of denaturation 30s, 56 DEG C of annealing 30s, 72 DEG C are prolonged
1min is stretched, 35 circulations are expanded；72 DEG C of extension 10min, 4 DEG C of preservations.
Reaction system are as follows: template DNA 30ng, primer Scaf4_1k11F 30ng, primer Scaf4_1k11R 30ng,
DNTPs 0.1 mmol/L, MgCl21 U of 2.5 mmol/L, Taq archaeal dna polymerase, 10 μ L of PCR buffer, uses ddH2O will be total
Volume is mended to 20 μ L.
The present invention has the advantages that method of the invention is somebody's turn to do for specifically identifying domestic agaricus bisporus main breed
Method time-consuming is short, easy to operate, high specificity, and it is accurate and time saving and energy saving to identify.Identified according to specific amplification products banding pattern,
It is strains A s2796 that product banding pattern, which contains only 1004bp band product, and containing only 801bp band product is bacterial strain W192, is contained
Having 1004bp and 801bp band product is bacterial strain good fortune mushroom 38, is bacterial strain containing 1004bp, 801bp and 252bp band product
A15.Using method of the invention, identifying domestic agaricus bisporus main breed only needs 1 day test period that can identify, and passes through
Conventional agronomy fruiting identification method needs time-consuming 45-60 days, and can not quick and precisely identify.
Detailed description of the invention
Fig. 1 is the SSR-PCR amplified production map of 4 domestic agaricus bisporus main breeds.Swimming lane 1-4 is to number in table 1
The bacterial strain of 1-4；M:DL10000 DNA Markers.
Specific embodiment
Embodiment 1
1, Taq archaeal dna polymerase, PCR buffer, MgCl2, dNTPs be purchased from Xiamen Thailand capital biology Co., Ltd, primer student on commission
The synthesis of work bioengineering (Shanghai) limited liability company, PCR product sequence is by Sangon Biotech (Shanghai) Co., Ltd.
Sequencing.
The core former times acid sequence of the special primer are as follows:
Scaf4_1k11F:5 '-CAATCACAATCCCTCCAAGC-3 ',
Scaf4_1k11R:5 '-AAACGAATTAGGAACGGTGG-3 '；
2,4 agaricus bisporus country main breeds are preserved in Edible Fungus Research Institute, Fujian Academy of Agricultural Sciences's genetic breeding research
Room, strain number and strain name are referring to (table one)
3, agaricus bisporus DNA extraction and electrophoresis
Agaricus bisporus DNA is extracted with DNA electrophoresis according to document [Chen Meiyuan, Liao Jianhua, Li Hongrong, Guo Zhongjie, Lu Zhenghui, Cai Dan
Phoenix, DNA fingerprint analysis [J] China's agronomy notification of the raw cultivation of agaricus bisporus bacterial strain genetic diversity of Wang Ze, 2009,
(04): 149-156.].
4, SSR-PCR reaction system and condition
Reaction system are as follows: template DNA 30ng, primer Scaf4_1k11F 30ng, primer Scaf4_1k11R 30ng, dNTPs
2.5 mmol/L of 0.1 mmol/L, MgCl2,1 U of Taq archaeal dna polymerase, PCR buffer 1 ×, total volume is mended with ddH2O
To 20 μ L.Amplification condition are as follows: 94 DEG C of initial denaturation 5min；94 DEG C of denaturation 30s, 56 DEG C of annealing 30s, 72 DEG C of extension 1min, amplification 35
A circulation；72 DEG C of extension 10min, 4 DEG C of preservations.PCR product electrophoresis according to document [Chen Meiyuan, Liao Jianhua, Wang Bo, Li Hongrong,
The DNA fingerprint analysis of the raw 90 bacterial strain genetic diversities of Chinese Wild Agaricus of Lu Zhenghui, Guo Zhongjie, Cai Danfeng, Wang Ze
[J] edible mushroom journal, 2009, (01): 11-16.].
5, interpretation of result
SSR-PCR testing result such as Fig. 1, No. 1-4 is 4 domestic agaricus bisporus main breed As2796, W192,38 and of good fortune mushroom
A15, it is strains A s2796 that wherein product banding pattern, which contains only 1004bp band product, and containing only 801bp band product is bacterium
Strain W192, is bacterial strain good fortune mushroom 38 containing 1004bp and 801bp band product, in product banding pattern containing 1004bp, 801bp and
252bp band product is strains A 15.Detect positive rate 100%.As it can be seen that this group of primer and its PCR product banding pattern of amplification can
Domestic 4 agaricus bisporus main breeds are efficiently differentiated and be identified.
4 strain numbers and strain name in table 1: Fig. 1
1 strains tested of table number and source
6, the sequence (1004bp) of specific PCR amplification product and primer position in the invention
5’-AAACGAATTAGGAACGGTGGTTGACAATGCAAGTTTAAATCGGATTTGGATTTTTTGCTGCGATGGAAG
GGGACTTGTACCATCCGAAGCGGCCCGGCTATTTCAAACAGATTAACGAATAATGTGCAGTGAATAGCATGTGAAA
AAGTAGATATCAATCAAAAGCAGATATATAAGTACAGGTAGGAATGACTGTAGTTTCACCAATAAGAAAATAAGTC
AGCTAAGGCTACAAATTGTCAATGGGATGGAGAGAGTGTCATACCGGATATATATACATAGTACTACACAGGATTG
ACCGAGGTTTGGGAAGATGGATATCGTTTTGACCTGCTCGATTCTATCCATGGGTCCGTCAGAGGTGAAGATAAGT
GACTACCAAATGACATGCAAAGGGTAAAGTCGACTATGATTGCAAAGCGAAGGAGCGAGGAGTAATGGAAAGAAAG
AAGGAAGAAGGAAGGAATCAAGGAATCAAGGAATCAAGGAAAGAAAGAAAGAAAGAAAGACAACAGACAACAGAAA
GCAAGAGGAGAAATACCGGTACGAGTGGGGACAGGATCAATTTCGTACACTTCACAGTCCCATCCTTTCTGCTAAC
GTCCGATCACCATTTATACCGTAGACATTGTTTGGTGTCGCATTGCTTGCTCCGTGGAATGCTTGGCCTCCTCCAG
CAGCTCTGTCTCCACGACGAGTGCGTCCAGGGCCGCCACCATTTCTCCTGCTGAGGTTGGTGTTCAAACCCTCTAT
CTCGACTGCGGTCCTGACCTTGAGGCCCTCTATCGCGATCCCGTTCTCGATTCCTCTCTCTCCCGCTCTCCTTATC
ATGTTCACGATCCATATCCTTCTCCCATCCGCGCCCAGATTCGGTGGGTAATGGGTGCTCATTATCAACGTCCATT
TTGGACGAGCGATTTCCTTTGTTGGCATCATGCCCTCCACGAACGTCTTTTGTCACTACGTTTTGGGTGATTGTCA
CCGCGCTTGGAGGATTGTGATTG-3’。
7, the sequence (801bp) of specific PCR amplification product and primer position in the invention
5’-AAACGAATTAGGAACGGTGGCCATGGAAGTCGGAACCCGCTAAGGAGTGTGTAACAACTCACCTGCCGA
ATGAACTAGCCCTGAAAATGGATGGCGCTTAAGCGTGATACCCATACCTCGCCGTCAGTGTTAAGGTGATGCGCTG
ACGAGTAGGCAGGCGTGGAGGTCAGTGAAGAAGCCTTGGCAGTGATGCTGGGTGAAACGGCCTCTAGTGCAGATCT
TGGTGGTAGTAGCAAATATTCAAGTGAGAACCTTGAAGACTGAAGTGGAGAAAGGTTCCATGGTAACAGCAGTTGG
ACATGGGTCAGTCGATCCTAAGAGATAGGGAAGCTCCGTTTCAAAGTGTGCGATTCTTCGCACCGCCTATCGAAAG
GGAATCCGGTTAAAATCCCGGAACCAGGATGTGGATCTTTAACGGCAACGTAACTGAACTTGGAGACGTTGGCAGG
GGCCCCGGGAAGAGTTATCTTTTCTCCTTAACAGTCTACCACCCTGAAATCAGATTGTCTGGAGCTAGGGTTCAAT
GACTGGTAGAGCACAACACCTCTGTTGTGTCCGGTGCGTTCCTGACAGCCCTTGAAAATCCAAGGGAATGGATAAT
TTTCACACCTGGTCGTACTCATAACCGCAGCAGGTCTCCAAGGTGAACAGCCTCTAGTTGATAGAACAATGTAGAT
AAGGGAAGTCGGCAAAATAGATCCGTAACTTCGGGAAAAGGATTGGCTCTAAGGGTTGGGGTGTGTCGGGCCTTGG
CTGGAAGCCTCGGGACCCAGCTGGGGACTGCTTGGAGGGATTGTGATT-3’。
8, the sequence (252bp) of specific PCR amplification product and primer position in the invention
5’-AAACGAATTAGGAACGGTGGTCCGCCTCCCCGCATTCATCAGCTACTTGAAGGACTTCCGGAACTCAGC
TCCCCCAGACAAGCAATAGAGAACCCTCATTTGCTGAAAGGCCGCAGCTTGTCTGACGGAACATTGATTTACACCC
CCCCCCCTGCTACGGTGGATCGAAGAGTGTAAACGAGGACAAATTGGGCGCACTCGAGTCCTCGGACGAGGAAGCG
CATTCATTGAAGGCTTGGAGGGATTGTGATT-3’。
The foregoing is merely presently preferred embodiments of the present invention, all equivalent changes done according to scope of the present invention patent with
Modification, is all covered by the present invention.
SEQUENCE LISTING
<110>Edible Fungus Research Institute, Fujian Academy of Agricultural Sciences (Fujian Mushroom Fungal Research Station)
<120>a kind of SSR-PCR identification method for distinguishing 4 main breeds of agaricus bisporus
<130> 5
<160> 5
<170> PatentIn version 3.3
<210> 1
<211> 20
<212> DNA
<213>artificial sequence
<400> 1
caatcacaat ccctccaagc 20
<210> 2
<211> 20
<212> DNA
<213>artificial sequence
<400> 2
aaacgaatta ggaacggtgg 20
<210> 3
<211> 1004
<212> DNA
<213>artificial sequence
<400> 3
aaacgaatta ggaacggtgg ttgacaatgc aagtttaaat cggatttgga ttttttgctg 60
cgatggaagg ggacttgtac catccgaagc ggcccggcta tttcaaacag attaacgaat 120
aatgtgcagt gaatagcatg tgaaaaagta gatatcaatc aaaagcagat atataagtac 180
aggtaggaat gactgtagtt tcaccaataa gaaaataagt cagctaaggc tacaaattgt 240
caatgggatg gagagagtgt cataccggat atatatacat agtactacac aggattgacc 300
gaggtttggg aagatggata tcgttttgac ctgctcgatt ctatccatgg gtccgtcaga 360
ggtgaagata agtgactacc aaatgacatg caaagggtaa agtcgactat gattgcaaag 420
cgaaggagcg aggagtaatg gaaagaaaga aggaagaagg aaggaatcaa ggaatcaagg 480
aatcaaggaa agaaagaaag aaagaaagac aacagacaac agaaagcaag aggagaaata 540
ccggtacgag tggggacagg atcaatttcg tacacttcac agtcccatcc tttctgctaa 600
cgtccgatca ccatttatac cgtagacatt gtttggtgtc gcattgcttg ctccgtggaa 660
tgcttggcct cctccagcag ctctgtctcc acgacgagtg cgtccagggc cgccaccatt 720
tctcctgctg aggttggtgt tcaaaccctc tatctcgact gcggtcctga ccttgaggcc 780
ctctatcgcg atcccgttct cgattcctct ctctcccgct ctccttatca tgttcacgat 840
ccatatcctt ctcccatccg cgcccagatt cggtgggtaa tgggtgctca ttatcaacgt 900
ccattttgga cgagcgattt cctttgttgg catcatgccc tccacgaacg tcttttgtca 960
ctacgttttg ggtgattgtc accgcgcttg gaggattgtg attg 1004
<210> 4
<211> 801
<212> DNA
<213>artificial sequence
<400> 4
aaacgaatta ggaacggtgg ccatggaagt cggaacccgc taaggagtgt gtaacaactc 60
acctgccgaa tgaactagcc ctgaaaatgg atggcgctta agcgtgatac ccatacctcg 120
ccgtcagtgt taaggtgatg cgctgacgag taggcaggcg tggaggtcag tgaagaagcc 180
ttggcagtga tgctgggtga aacggcctct agtgcagatc ttggtggtag tagcaaatat 240
tcaagtgaga accttgaaga ctgaagtgga gaaaggttcc atggtaacag cagttggaca 300
tgggtcagtc gatcctaaga gatagggaag ctccgtttca aagtgtgcga ttcttcgcac 360
cgcctatcga aagggaatcc ggttaaaatc ccggaaccag gatgtggatc tttaacggca 420
acgtaactga acttggagac gttggcaggg gccccgggaa gagttatctt ttctccttaa 480
cagtctacca ccctgaaatc agattgtctg gagctagggt tcaatgactg gtagagcaca 540
acacctctgt tgtgtccggt gcgttcctga cagcccttga aaatccaagg gaatggataa 600
ttttcacacc tggtcgtact cataaccgca gcaggtctcc aaggtgaaca gcctctagtt 660
gatagaacaa tgtagataag ggaagtcggc aaaatagatc cgtaacttcg ggaaaaggat 720
tggctctaag ggttggggtg tgtcgggcct tggctggaag cctcgggacc cagctgggga 780
ctgcttggag ggattgtgat t 801
<210> 5
<211> 252
<212> DNA
<213>artificial sequence
<400> 5
aaacgaatta ggaacggtgg tccgcctccc cgcattcatc agctacttga aggacttccg 60
gaactcagct cccccagaca agcaatagag aaccctcatt tgctgaaagg ccgcagcttg 120
tctgacggaa cattgattta cacccccccc cctgctacgg tggatcgaag agtgtaaacg 180
aggacaaatt gggcgcactc gagtcctcgg acgaggaagc gcattcattg aaggcttgga 240
gggattgtga tt 252
Claims (5)
1. a kind of SSR-PCR for distinguishing 4 main breeds of agaricus bisporus identifies primer, it is characterised in that: the primer sequence are as follows:
Scaf4_1k11F:5 '-CAATCACAATCCCTCCAAGC-3 ',
Scaf4_1k11R:5 '-AAACGAATTAGGAACGGTGG-3 '.
2. a kind of SSR-PCR for distinguishing 4 main breeds of agaricus bisporus according to claim 1 identifies primer, feature
Be: 4 main breeds of the agaricus bisporus are kind As2796, W192, good fortune mushroom 38 and A15.
3. it is used to distinguish the SSR-PCR identification method of 4 main breeds of agaricus bisporus using primer described in claim 1, it is special
Sign is: using the primer pair Scaf4_1k11F and Scaf4_1k11R described in claim 1 to the main cultivation of agaricus bisporus
The genomic DNA of kind As2796, W192, good fortune mushroom 38 and A15 carry out SSR-PCR amplification, specifically AFLP system are as follows:
As2796 amplifies the product of 1004bp；W192 amplifies the product of 801bp；Good fortune mushroom 38 amplifies the production of 1004bp and 801bp
Object；A15 amplifies the product of 1004bp, 801bp and 252bp.
4. a kind of SSR-PCR identification method for distinguishing 4 main breeds of agaricus bisporus according to claim 3, feature
It is: the condition of the SSR-PCR amplification are as follows: 94 DEG C of initial denaturation 5min；94 DEG C of denaturation 30s, 56 DEG C of annealing 30s, 72 DEG C extend
1min expands 35 circulations；72 DEG C of extension 10min, 4 DEG C of preservations.
5. a kind of SSR-PCR identification method for distinguishing 4 main breeds of agaricus bisporus according to claim 3, feature
It is: the reaction system of the SSR-PCR amplification are as follows: template DNA 30ng, primer Scaf4_1k11F 30ng, primer Scaf4_
1k11R 30ng, dNTPs 0.1 mmol/L, MgCl21 U of 2.5 mmol/L, Taq archaeal dna polymerase, 1 × PCR buffer 10
μ L, uses ddH2O mends total volume to 20 μ L.
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|---|---|---|---|---|
| CN111235294A (en) * | 2020-02-21 | 2020-06-05 | 拉萨市高原生物研究所 | DNA bar code and primer for screening high-quality Tibetan brown mushroom and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105200133A (en) * | 2015-09-24 | 2015-12-30 | 福建省农业科学院食用菌研究所 | ISSR marker for authenticating high-quality agaricus bisporus bacterial strain and application |
| CN105255882A (en) * | 2015-11-22 | 2016-01-20 | 吉林农业大学 | Agaricus bisporus ssr molecular marker specific primer system and application thereof |
| WO2018013603A1 (en) * | 2016-07-12 | 2018-01-18 | Amycel, Inc. | White hybrid mushrooms for commercial production |
-
2018
- 2018-10-29 CN CN201811269851.0A patent/CN109207625B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105200133A (en) * | 2015-09-24 | 2015-12-30 | 福建省农业科学院食用菌研究所 | ISSR marker for authenticating high-quality agaricus bisporus bacterial strain and application |
| CN105255882A (en) * | 2015-11-22 | 2016-01-20 | 吉林农业大学 | Agaricus bisporus ssr molecular marker specific primer system and application thereof |
| WO2018013603A1 (en) * | 2016-07-12 | 2018-01-18 | Amycel, Inc. | White hybrid mushrooms for commercial production |
Non-Patent Citations (4)
| Title |
|---|
| MARIE FOULONGNE-ORIOL等: "Novel microsatellite markers suitable for genetic studies in the white button mushroom Agaricus bisporus", 《APPLIED MICROBIOLOGY AND BIOTECHNOLOGY》 * |
| 王新新等: "双孢蘑菇种质SSR分子身份证的构建", 《食用菌学报》 * |
| 王泽生等: "利用SRAP和ISSR分子标记技术分析双孢蘑菇的遗传多样性", 《2012年中国菌物学会学术年会会议摘要》 * |
| 顾敏等: "双孢蘑菇 SSＲ 分子标记开发及其在遗传多样性分析中的应用", 《浙江农业学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111235294A (en) * | 2020-02-21 | 2020-06-05 | 拉萨市高原生物研究所 | DNA bar code and primer for screening high-quality Tibetan brown mushroom and application thereof |
| CN111235294B (en) * | 2020-02-21 | 2023-07-07 | 拉萨市高原生物研究所 | DNA bar code and primer for screening high-quality Tibetan brown mushrooms and application of DNA bar code and primer |
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