基于多重PCR-LDR技术建立近交系大鼠单核苷酸多态性遗传检测方案

doi:10.12300/j.issn.1674-5817.2023.073

实验动物与比较医学 ›› 2023, Vol. 43 ›› Issue (5): 548-558.DOI: 10.12300/j.issn.1674-5817.2023.073

基于多重PCR-LDR技术建立近交系大鼠单核苷酸多态性遗传检测方案

赵丽亚¹^,²(), 倪丽菊¹(), 张彩勤³, 汤建平¹^,², 姚养正⁴, 聂艳艳¹, 顾晓雪², 赵莹¹()()

^1.上海实验动物研究中心, 上海 201203
^2.上海必凯科翼生物科技有限公司, 上海 201203
^3.空军军医大学实验动物中心, 西安 710038
^4.陕西省中医药研究院, 西安 710003

收稿日期:2023-06-09 修回日期:2023-08-08 出版日期:2023-10-25 发布日期:2023-11-01
通讯作者: 赵莹（1982—），女，硕士，副研究员，研究方向：实验动物遗传。E-mail： zhaoying@slarc.org.cn。ORCID： 0000-0002-0551-0566
作者简介:赵丽亚（1981—），女，硕士，工程师，研究方向：分子遗传学。E-mail： Zhaoliya3002@163.com；
倪丽菊（1979—），女，硕士，副研究员，研究方向：实验动物遗传。E-mail： niliju@163.com

Establishing a Genetic Detection Protocol of Single Nucleotide Polymorphisms Panels in Inbred Rats Based on Multiplex PCR-LDR

Liya ZHAO¹^,²(), Liju NI¹(), Caiqin ZHANG³, Jianping TANG¹^,², Yangzheng YAO⁴, Yanyan NIE¹, Xiaoxue GU², Ying ZHAO¹()()

^1.Shanghai Laboratory Animal Research Center, Shanghai 201203, China
^2.Shanghai BK/KY Biotechnology Co. , Ltd. , Shanghai 201203, China
^3.Laboratory Animal Center of Air Force Medical University, Xi'an 710038, China
^4.Shaanxi Academy of Traditional Chinese Medicine, Xi'an 710003, China

Received:2023-06-09 Revised:2023-08-08 Published:2023-10-25 Online:2023-11-01
Contact: ZHAO Ying (ORCID: 0000-0002-0551-0566), E-mail: zhaoying@slarc.org.cn

摘要/Abstract

摘要：

目的建立一套基于多重PCR-连接酶检测反应（ligase detection reaction，LDR）技术的近交系大鼠单核苷酸多态性（single nucleotide polymorphisms，SNP）检测方案。方法在5个品系的SPF级近交系大鼠1～20号常染色体和X染色体上共选取40个大鼠SNP位点，将SNP位点随机分为4组，构建基于多重PCR-LDR技术的近交系大鼠4组SNP位点基因检测方案。采用本方案检测国内另两家大鼠供应商的9个常用大鼠品系。最后，通过第三方实验室对不同DNA聚合酶的扩增效果进行比对，验证本方案的可行性。结果用所构建的近交系大鼠SNP遗传检测方案测试5个大鼠品系时，各样本的所有位点均得到了良好的扩增结果。采用本方案检测国内另两家大鼠供应商的9个常用大鼠品系时也得到了良好的扩增结果，40个SNP位点在每个近交系大鼠中均为纯合。用3种来源不同的DNA聚合酶同时检测相同大鼠DNA样本的结果显示，Multiplex PCR Kit、AmpliTaq Gold? 360 DNA聚合酶、PlatinumⅡ Taq热启动DNA聚合酶在第1～3组SNP位点均有扩增产物的电泳峰，其中PlatinumⅡ Taq热启动DNA聚合酶在第4组SNP位点中少了一个扩增产物的电泳峰。另外，不同实验室间的比对结果显示，相同扩增体系的检测结果一致。结论基于多重PCR-LDR技术成功建立了一套覆盖所有常染色体与X染色体的大鼠SNP检测方案，该方法的稳定性和重复性俱佳。

关键词: 近交系大鼠, 单核苷酸多态性, 多重PCR-LDR, 遗传检测, 验证

Abstract:

Objective To establish a set of single nucleotide polymorphisms (SNP) detection protocol for inbred rats based on multiplex PCR-ligase detection reaction (LDR). Methods A total of 40 rats SNP sites were selected on chromosomes 1-20 and X of rats among 5 inbred strains of rats, and the 40 SNP sites were randomly divided into four groups. A genetic detection protocol for 4 groups of SNP in inbred rats based on multiplex PCR-LDR technology was constructed. 9 commonly used rat strains from two other domestic rat suppliers were detected by this protocol. Finally, the feasibility of this protocol was verified by comparing the amplification effects of different DNA polymerases by a third-party laboratory. Results When using the constructed SNP detection protocol for inbred rats to test 5 rat strains, all sites in each sample obtained good amplification results. The 9 commonly used rat strains from two other rat suppliers in china were also well amplified by this SNP detection protocol, and 40 SNPs were homozygous in each Inbred strain. The results of detection of the same rat DNA samples with three different DNA polymerases showed that the Multiplex PCR Kit, AmpliTaq Gold 360 DNA polymerase and Platinum II Taq hot start DNA polymerase had electrophoretic peaks of amplification products at all SNP sites in groups 1 to 3, and Platinum II Taq hot start DNA polymerase had one less electrophoretic peak of the amplification products at the SNP sites in group 4. In addition, inter-laboratory comparisons showed consistent results for the same amplification system. Conclusion Based on multiplex PCR-LDR technology, this study successfully established a SNP detection protocol for rats covering all autosomes and X chromosomes with the excellent stability and repeatability.

Key words: Inbred rats, Single Nucleotide Polymorphisms, Multiplex PCR-LDR, Genetic detection, Verification

中图分类号:

Q95-33

赵丽亚, 倪丽菊, 张彩勤, 汤建平, 姚养正, 聂艳艳, 顾晓雪, 赵莹. 基于多重PCR-LDR技术建立近交系大鼠单核苷酸多态性遗传检测方案[J]. 实验动物与比较医学, 2023, 43(5): 548-558.

Liya ZHAO, Liju NI, Caiqin ZHANG, Jianping TANG, Yangzheng YAO, Yanyan NIE, Xiaoxue GU, Ying ZHAO. Establishing a Genetic Detection Protocol of Single Nucleotide Polymorphisms Panels in Inbred Rats Based on Multiplex PCR-LDR[J]. Laboratory Animal and Comparative Medicine, 2023, 43(5): 548-558.

图/表 6

参考文献 13

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序号 No.	SNP位点 Reference SNP ID	染色体 Chromosome	引物名称 Primer name	引物序列 Primer sequences
a1	rs65970293	chr1	rs65970293-up	TGCACATTGGGACTGCTTTA
a1	rs65970293	chr1	rs65970293-low	ATCTGGCATGGACAGAGCTT
a2	rs106996961	chr2	rs106996961-up	ATAGGGCCAAGGTCCCATTC
a2	rs106996961	chr2	rs106996961-low	GAAAAGAACGCTAATCACAGGACA
a3	rs63860341	chr3	rs63860341-up	CAATGAATCAAAGGCAACCA
a3	rs63860341	chr3	rs63860341-low	CCACGGTCCCTATGTGAACT
a4	rs64750237	chr4	rs64750237-up	CAGGTATGGCTTTGGAGGAA
a4	rs64750237	chr4	rs64750237-low	GTTTTCAGCTGGAGCTGTCC
a5	rs107261116	chr5	rs107261116-up	CAGCATTGGCAAGTCTTCCT
a5	rs107261116	chr5	rs107261116-low	TGATAAAAGGAGCAGCAGCA
a6	rs64102787	chr6	rs64102787-up	CGGGGATCAAAGAGACAAAA
a6	rs64102787	chr6	rs64102787-low	ATAGTGCCAAAGCCCAACAC
a7	rs105187832	chr7	rs105187832-up	AGACTACTGGGCTGCTCACC
a7	rs105187832	chr7	rs105187832-low	CATGGCACACCTTTCTTTCA
a8	rs64682441	chr9	rs64682441-up	GCTGGAAGTCTCTGGAGTGG
a8	rs64682441	chr9	rs64682441-low	ACAAGTGGACTGGCTCAGGT
a9	rs65067166	chr10	rs65067166-up	ATCCCTCTCTTTGCCAGGAT
a9	rs65067166	chr10	rs65067166-low	GTAGGGTGAGGCTGCTTCTG
a10	rs8167985	chr11	rs8167985-up	ACCCCCTCTTGCTTTCTGTT
a10	rs8167985	chr11	rs8167985-low	CTGCTCTTGGATGAAAACCA
b1	rs106672258	chr12	rs106672258-up	GCCCTTCCCAAGGTCATACT
b1	rs106672258	chr12	rs106672258-low	AAAGCACAAAAATGCCATCC
b2	rs65921305	chr13	rs65921305-up	AACCTGGTGATTTGTCAGAGC
b2	rs65921305	chr13	rs65921305-low	GGCAGACTCCAGGAAGATTG
b3	rs106458396	chr15	rs106458396-up	AGAGCAGCTGCCAAAATAGC
b3	rs106458396	chr15	rs106458396-low	AAACCTGGCTTTAGCAAGCA
b4	rs106888236	chr16	rs106888236-up	TGTGAAGAACACAGGCTCTCC
b4	rs106888236	chr16	rs106888236-low	TGCCAAACACCATGTAATGAA
b5	rs63814610	chr17	rs63814610-up	TAGGACAGGGGGAACAACTG
b5	rs63814610	chr17	rs63814610-low	CCCAAGCATCCTTGACTGTT
b6	rs8152541	chr18	rs8152541-up	CGTGTCATTCCTGACTGTGG
b6	rs8152541	chr18	rs8152541-low	ACAGGAGCTGGCACTGAATC
b7	rs105824121	chr19	rs105824121-up	TTCTGCAATCAAGGGTGGAT
b7	rs105824121	chr19	rs105824121-low	GGCCACCAAAGATACACAGG
b8	rs13453252	chr20	rs13453252-up	TACGCCATTTTGCGACATTA
b8	rs13453252	chr20	rs13453252-low	GGTTCCCTACGAGGAACGAC
b9	rs65108991	chrX	rs65108991-up	CATCCCAGAAGCCCTGTTTA
b9	rs65108991	chrX	rs65108991-low	GAAGTCAGTGCTGTGCCTCA
c1	rs106891788	chr1	rs106891788-up	TCCATCTGCTTGGCCCTAAT
c1	rs106891788	chr1	rs106891788-low	GTTGCAACCCACAGGTTGAG
c2	rs107493604	chr2	rs107493604-up	TCCTCCTGGAAGACAGCAAC
c2	rs107493604	chr2	rs107493604-low	GTTCCTGAGAAGGGGCTCTC
c3	rs65632519	chr3	rs65632519-up	CCCTGTTTCAGCCTTTTGTG
c3	rs65632519	chr3	rs65632519-low	GAGGAAGAGGGAAGGGAGAA
c4	rs107163178	chr4	rs107163178-up	TTCTCCCACCGGAGACTTAG
c4	rs107163178	chr4	rs107163178-low	GAGAGGGCATATGTGGGATG
c5	rs105170847	chr5	rs105170847-up	TGGTTGGATGAATGGAGACA
c5	rs105170847	chr5	rs105170847-low	CCTCAGGAGCACCAACATTT
c6	rs106547562	chr6	rs106547562-up	GGTTTTGTCCTCAGCAGGTT
c6	rs106547562	chr6	rs106547562-low	CTCTTAGGCATCTGGGGAAA
c7	rs65502317	chr7	rs65502317-up	TGTGCATGCTTACAGTGGCTA
c7	rs65502317	chr7	rs65502317-low	AGCCAACTGGAGCCACATAG
c8	rs105694545	chr8	rs105694545-up	TCAGCAGTGCTCAGCAGTTT
c8	rs105694545	chr8	rs105694545-low	CGTAACATCATTTGGCATCG
c9	rs65801920	chr9	rs65801920-up	TGGCTCTTGGTCATCATCTG
c9	rs65801920	chr9	rs65801920-low	TTTTGCAGCCTGTGTGTAGC
c10	rs105168947	chr10	rs105168947-up	GGCGGACAGAACATAGGTGT
c10	rs105168947	chr10	rs105168947-low	CAGCCAAACTCCCAGATCAT
c11	rs106880606	chr11	rs106880606-up	AGTGGGTTTAGCAAGCAGGA
c11	rs106880606	chr11	rs106880606-low	GTCGTCGAAATGCAGGAGAT
d1	rs8158863	chr12	rs8158863-up	GGTCACAGCCATTCTGGATT
d1	rs8158863	chr12	rs8158863-low	GCCCTGAATGCAGGTGTAGT
d2	rs105965886	chr13	rs105965886-up	CACCAGCAAAAGGGAAAAAG
d2	rs105965886	chr13	rs105965886-low	CAGAACACAGCCTCAGTTGG
d3	rs63919332	chr14	rs63919332-up	GTCCTGTCGCCCAATATCAC
d3	rs63919332	chr14	rs63919332-low	ACAACAATGGCAGTGGTCAA
d4	rs65809306	chr15	rs65809306-up	AGCACTTGTGTGGGAAGCTC
d4	rs65809306	chr15	rs65809306-low	CCTGACAAAACCACATGCAG
d5	rs65087257	chr16	rs65087257-up	CCTTCCTTTTCCACTGTCCA
d5	rs65087257	chr16	rs65087257-low	CTTAGGGCCTAGGGGCTTTA
d6	rs106021355	chr17	rs106021355-up	GGTGGTTTATGGCCAGGATA
d6	rs106021355	chr17	rs106021355-low	AACTGGAGAGGAAAAGGGACA
d7	rs106171441	chr18	rs106171441-up	TCAATTCTTGGCAGCATCAG
d7	rs106171441	chr18	rs106171441-low	CTGGCCACTTCTCTGAGTCC
d8	rs106777713	chr19	rs106777713-up	GTGAAGCGACATCGTCAAGA
d8	rs106777713	chr19	rs106777713-low	CTTGGCTGTGAGATCGAGGT
d9	rs104937988	chr20	rs104937988-up	GCTGCACAGAACTGATTCCA
d9	rs104937988	chr20	rs104937988-low	TGCAAGAATTAAATAAAATCCAAA
d10	rs105294203	chrX	rs105294203-up	ACTCCCCAGAGGATTGTCG
d10	rs105294203	chrX	rs105294203-low	ATGTTCAAGTGCAACGATGG

序号 No.	SNP位点 Reference SNP ID	染色体 Chromosome	引物名称 Primer name	引物序列 Primer sequences
a1	rs65970293	chr1	rs65970293-up	TGCACATTGGGACTGCTTTA
a1	rs65970293	chr1	rs65970293-low	ATCTGGCATGGACAGAGCTT
a2	rs106996961	chr2	rs106996961-up	ATAGGGCCAAGGTCCCATTC
a2	rs106996961	chr2	rs106996961-low	GAAAAGAACGCTAATCACAGGACA
a3	rs63860341	chr3	rs63860341-up	CAATGAATCAAAGGCAACCA
a3	rs63860341	chr3	rs63860341-low	CCACGGTCCCTATGTGAACT
a4	rs64750237	chr4	rs64750237-up	CAGGTATGGCTTTGGAGGAA
a4	rs64750237	chr4	rs64750237-low	GTTTTCAGCTGGAGCTGTCC
a5	rs107261116	chr5	rs107261116-up	CAGCATTGGCAAGTCTTCCT
a5	rs107261116	chr5	rs107261116-low	TGATAAAAGGAGCAGCAGCA
a6	rs64102787	chr6	rs64102787-up	CGGGGATCAAAGAGACAAAA
a6	rs64102787	chr6	rs64102787-low	ATAGTGCCAAAGCCCAACAC
a7	rs105187832	chr7	rs105187832-up	AGACTACTGGGCTGCTCACC
a7	rs105187832	chr7	rs105187832-low	CATGGCACACCTTTCTTTCA
a8	rs64682441	chr9	rs64682441-up	GCTGGAAGTCTCTGGAGTGG
a8	rs64682441	chr9	rs64682441-low	ACAAGTGGACTGGCTCAGGT
a9	rs65067166	chr10	rs65067166-up	ATCCCTCTCTTTGCCAGGAT
a9	rs65067166	chr10	rs65067166-low	GTAGGGTGAGGCTGCTTCTG
a10	rs8167985	chr11	rs8167985-up	ACCCCCTCTTGCTTTCTGTT
a10	rs8167985	chr11	rs8167985-low	CTGCTCTTGGATGAAAACCA
b1	rs106672258	chr12	rs106672258-up	GCCCTTCCCAAGGTCATACT
b1	rs106672258	chr12	rs106672258-low	AAAGCACAAAAATGCCATCC
b2	rs65921305	chr13	rs65921305-up	AACCTGGTGATTTGTCAGAGC
b2	rs65921305	chr13	rs65921305-low	GGCAGACTCCAGGAAGATTG
b3	rs106458396	chr15	rs106458396-up	AGAGCAGCTGCCAAAATAGC
b3	rs106458396	chr15	rs106458396-low	AAACCTGGCTTTAGCAAGCA
b4	rs106888236	chr16	rs106888236-up	TGTGAAGAACACAGGCTCTCC
b4	rs106888236	chr16	rs106888236-low	TGCCAAACACCATGTAATGAA
b5	rs63814610	chr17	rs63814610-up	TAGGACAGGGGGAACAACTG
b5	rs63814610	chr17	rs63814610-low	CCCAAGCATCCTTGACTGTT
b6	rs8152541	chr18	rs8152541-up	CGTGTCATTCCTGACTGTGG
b6	rs8152541	chr18	rs8152541-low	ACAGGAGCTGGCACTGAATC
b7	rs105824121	chr19	rs105824121-up	TTCTGCAATCAAGGGTGGAT
b7	rs105824121	chr19	rs105824121-low	GGCCACCAAAGATACACAGG
b8	rs13453252	chr20	rs13453252-up	TACGCCATTTTGCGACATTA
b8	rs13453252	chr20	rs13453252-low	GGTTCCCTACGAGGAACGAC
b9	rs65108991	chrX	rs65108991-up	CATCCCAGAAGCCCTGTTTA
b9	rs65108991	chrX	rs65108991-low	GAAGTCAGTGCTGTGCCTCA
c1	rs106891788	chr1	rs106891788-up	TCCATCTGCTTGGCCCTAAT
c1	rs106891788	chr1	rs106891788-low	GTTGCAACCCACAGGTTGAG
c2	rs107493604	chr2	rs107493604-up	TCCTCCTGGAAGACAGCAAC
c2	rs107493604	chr2	rs107493604-low	GTTCCTGAGAAGGGGCTCTC
c3	rs65632519	chr3	rs65632519-up	CCCTGTTTCAGCCTTTTGTG
c3	rs65632519	chr3	rs65632519-low	GAGGAAGAGGGAAGGGAGAA
c4	rs107163178	chr4	rs107163178-up	TTCTCCCACCGGAGACTTAG
c4	rs107163178	chr4	rs107163178-low	GAGAGGGCATATGTGGGATG
c5	rs105170847	chr5	rs105170847-up	TGGTTGGATGAATGGAGACA
c5	rs105170847	chr5	rs105170847-low	CCTCAGGAGCACCAACATTT
c6	rs106547562	chr6	rs106547562-up	GGTTTTGTCCTCAGCAGGTT
c6	rs106547562	chr6	rs106547562-low	CTCTTAGGCATCTGGGGAAA
c7	rs65502317	chr7	rs65502317-up	TGTGCATGCTTACAGTGGCTA
c7	rs65502317	chr7	rs65502317-low	AGCCAACTGGAGCCACATAG
c8	rs105694545	chr8	rs105694545-up	TCAGCAGTGCTCAGCAGTTT
c8	rs105694545	chr8	rs105694545-low	CGTAACATCATTTGGCATCG
c9	rs65801920	chr9	rs65801920-up	TGGCTCTTGGTCATCATCTG
c9	rs65801920	chr9	rs65801920-low	TTTTGCAGCCTGTGTGTAGC
c10	rs105168947	chr10	rs105168947-up	GGCGGACAGAACATAGGTGT
c10	rs105168947	chr10	rs105168947-low	CAGCCAAACTCCCAGATCAT
c11	rs106880606	chr11	rs106880606-up	AGTGGGTTTAGCAAGCAGGA
c11	rs106880606	chr11	rs106880606-low	GTCGTCGAAATGCAGGAGAT
d1	rs8158863	chr12	rs8158863-up	GGTCACAGCCATTCTGGATT
d1	rs8158863	chr12	rs8158863-low	GCCCTGAATGCAGGTGTAGT
d2	rs105965886	chr13	rs105965886-up	CACCAGCAAAAGGGAAAAAG
d2	rs105965886	chr13	rs105965886-low	CAGAACACAGCCTCAGTTGG
d3	rs63919332	chr14	rs63919332-up	GTCCTGTCGCCCAATATCAC
d3	rs63919332	chr14	rs63919332-low	ACAACAATGGCAGTGGTCAA
d4	rs65809306	chr15	rs65809306-up	AGCACTTGTGTGGGAAGCTC
d4	rs65809306	chr15	rs65809306-low	CCTGACAAAACCACATGCAG
d5	rs65087257	chr16	rs65087257-up	CCTTCCTTTTCCACTGTCCA
d5	rs65087257	chr16	rs65087257-low	CTTAGGGCCTAGGGGCTTTA
d6	rs106021355	chr17	rs106021355-up	GGTGGTTTATGGCCAGGATA
d6	rs106021355	chr17	rs106021355-low	AACTGGAGAGGAAAAGGGACA
d7	rs106171441	chr18	rs106171441-up	TCAATTCTTGGCAGCATCAG
d7	rs106171441	chr18	rs106171441-low	CTGGCCACTTCTCTGAGTCC
d8	rs106777713	chr19	rs106777713-up	GTGAAGCGACATCGTCAAGA
d8	rs106777713	chr19	rs106777713-low	CTTGGCTGTGAGATCGAGGT
d9	rs104937988	chr20	rs104937988-up	GCTGCACAGAACTGATTCCA
d9	rs104937988	chr20	rs104937988-low	TGCAAGAATTAAATAAAATCCAAA
d10	rs105294203	chrX	rs105294203-up	ACTCCCCAGAGGATTGTCG
d10	rs105294203	chrX	rs105294203-low	ATGTTCAAGTGCAACGATGG

大鼠品系 Rat strains	碱基有差异的SNP位点编号（右上三角）及数量（左下三角） Differences in SNP loci (right-upper triangle) and numbers (left-lower triangle)
大鼠品系 Rat strains	F344/BKl	BN/BKl	DA/BKl	PVG/BKl	Lewis/BKl
F344/BKl	-	a1/a2/a3/a4/a5/a9/a10/b2/b5/b7/c2/c3/c6/c8/c10/c11/d2/d6/d7/d8/d9	a1/a2/a4/a7/a9/b4/b8/c1/c2/c3/c6/c9/c11/d2/d7/d8/d10	a4/b1/b3/b4/b5/b6/b7/c1/c6/c8/c9/d1/d2	a2/a4/a7/b2/b7/c1/c2/c4/c5/c7/c8/c11/d1/d2/d7/d8
BN/BKl	21	-	a3/a5/a7/a10/b2/b4/b5/b7/b8/c1/c8/c9/c10/d6/d9/d10	a1/a2/a3/a5/a9/a10/b1/b2/b3/b4/b6/c1/c2/c3/c9/c10/c11/d1/d6/d7/d8/d9	a1/a3/a5/a7/a9/a10/b5/c1/c3/c4/c5/c6/c7/c10/d1/d6/d9
DA/BKl	17	16	-	a1/a2/a7/a9/b1/b3/b5/b6/b7/b8/c2/c3/c8/c11/d1/d7/d8/d10	a1/a9/b2/b4/b7/b8/c3/c4/c5/c6/c7/c8/c9/d1/d10
PVG/BKl	13	22	18	-	a2/a7/b1/b2/b3/b4/b5/b6/c2/c4/c5/c6/c7/c9/c11/d7/d8
Lewis/BKl	16	17	15	17	-

大鼠品系 Rat strains	碱基有差异的SNP位点编号（右上三角）及数量（左下三角） Differences in SNP loci (right-upper triangle) and numbers (left-lower triangle)
大鼠品系 Rat strains	F344/BKl	BN/BKl	DA/BKl	PVG/BKl	Lewis/BKl
F344/BKl	-	a1/a2/a3/a4/a5/a9/a10/b2/b5/b7/c2/c3/c6/c8/c10/c11/d2/d6/d7/d8/d9	a1/a2/a4/a7/a9/b4/b8/c1/c2/c3/c6/c9/c11/d2/d7/d8/d10	a4/b1/b3/b4/b5/b6/b7/c1/c6/c8/c9/d1/d2	a2/a4/a7/b2/b7/c1/c2/c4/c5/c7/c8/c11/d1/d2/d7/d8
BN/BKl	21	-	a3/a5/a7/a10/b2/b4/b5/b7/b8/c1/c8/c9/c10/d6/d9/d10	a1/a2/a3/a5/a9/a10/b1/b2/b3/b4/b6/c1/c2/c3/c9/c10/c11/d1/d6/d7/d8/d9	a1/a3/a5/a7/a9/a10/b5/c1/c3/c4/c5/c6/c7/c10/d1/d6/d9
DA/BKl	17	16	-	a1/a2/a7/a9/b1/b3/b5/b6/b7/b8/c2/c3/c8/c11/d1/d7/d8/d10	a1/a9/b2/b4/b7/b8/c3/c4/c5/c6/c7/c8/c9/d1/d10
PVG/BKl	13	22	18	-	a2/a7/b1/b2/b3/b4/b5/b6/c2/c4/c5/c6/c7/c9/c11/d7/d8
Lewis/BKl	16	17	15	17	-

序号 No.	SNP标记名称 Reference SNP ID	染色体 Chromosome	不同品系大鼠的基因型 Genotype of different strains of rats
序号 No.	SNP标记名称 Reference SNP ID	染色体 Chromosome	F344/BKl	BN/BKl	DA/BKl	PVG/BKl	Lewis/BKl	SHR/Slac	GK/Slac	F344/JclSlac	Wistar/Slac	WKY/Slac	CXB	CHN1	CHN2	CHN5
a1	rs65970293	chr1	C/C	T/T	T/T	C/C	C/C	T/T	C/C	C/C	C/C	T/T	T/T	T/T	T/T	T/T
a2	rs106996961	chr2	T/T	G/G	G/G	T/T	G/G	G/G	G/G	T/T	T/T	G/G	G/G	G/G	G/G	T/T
a3	rs63860341	chr3	T/T	C/C	T/T	T/T	T/T	T/T	T/T	T/T	T/T	T/T	T/T	T/T	T/T	T/T
a4	rs64750237	chr4	C/C	T/T	T/T	T/T	T/T	C/C	C/C	C/C	T/T	C/C	T/T	T/T	T/T	T/T
a5	rs107261116	chr5	C/C	G/G	C/C	C/C	C/C	G/G	C/C	C/C	C/C	C/C	C/C	C/C	G/G	G/G
a6	rs64102787	chr6	C/C	C/C	C/C	C/C	C/C	A/A	A/A	C/C	C/C	A/A	C/C	C/C	C/C	A/A
a7	rs105187832	chr7	G/G	G/G	T/T	G/G	T/T	G/G	T/T	G/G	T/T	T/T	G/G	G/G	G/G	G/G
a8	rs64682441	chr9	T/T	T/T	T/T	T/T	T/T	C/C	C/C	T/T	T/T	C/C	C/C	C/C	C/C	C/C
a9	rs65067166	chr10	C/C	G/G	G/G	C/C	C/C	C/C	C/C	C/C	C/C	C/C	G/G	G/G	G/G	G/G
a10	rs8167985	chr11	G/G	T/T	G/G	G/G	G/G	T/T	G/G	G/G	G/G	G/G	G/G	G/G	G/G	G/G
b1	rs106672258	chr12	T/T	T/T	T/T	C/C	T/T	C/C	C/C	T/T	T/T	C/C	C/C	C/C	C/C	C/C
b2	rs65921305	chr13	G/G	T/T	G/G	G/G	T/T	G/G	G/G	G/G	T/T	G/G	G/G	G/G	G/G	T/T
b3	rs106458396	chr15	T/T	T/T	T/T	C/C	T/T	C/C	T/T	T/T	C/C	T/T	T/T	T/T	T/T	T/T
b4	rs106888236	chr16	C/C	C/C	A/A	A/A	C/C	A/A	A/A	C/C	A/A	C/C	C/C	A/A	A/A	C/C
b5	rs63814610	chr17	T/T	C/C	T/T	C/C	T/T	T/T	T/T	T/T	C/C	T/T	C	T/T	T/T	T/T
b6	rs8152541	chr18	G/G	G/G	G/G	A/A	G/G	A/A	A/A	G/G	G/G	G/G	A/A	A/A	G/G	A/A
b7	rs105824121	chr19	G/G	A/A	G/G	A/A	A/A	G/G	G/G	G/G	G/G	G/G	A/A	A/A	A/A	G/G
b8	rs13453252	chr20	G/G	G/G	C/C	G/G	G/G	G/G	C/C	G/G	C/C	C/C	G/G	G/G	G/G	G/G
b9	rs65108991	chrX	A/A	A/A	A/A	A/A	A/A	C/C	A/A	A/A	A/A	C/C	A/A	A/A	A/A	A/A
c1	rs106891788	chr1	C/C	C/C	T/T	T/T	T/T	T/T	C/C	C/C	T/T	C/C	T/T	T/T	T/T	T/T
c2	rs107493604	chr2	A/A	G/G	G/G	A/A	G/G	G/G	A/A	A/A	A/A	G/G	G/G	G/G	G/G	G/G
c3	rs65632519	chr3	T/T	A/A	A/A	T/T	T/T	A/A	A/A	T/T	T/T	T/T	A	T/T	T/T	T/T
c4	rs17163178	chr4	C/C	C/C	C/C	C/C	T/T	T/T	C/C	C/C	T/T	C/C	C/C	T/T	T/T	C/C
c5	rs105170847	chr5	C/C	C/C	C/C	C/C	T/T	C/C	C/C	C/C	C/C	C/C	C/C	C/C	C/C	C/C
c6	rs106547562	chr6	A/A	C/C	C/C	C/C	A/A	C/C	C/C	A/A	A/A	C/C	A/A	A/A	A/A	C/C
c7	rs65502317	chr7	C/C	C/C	C/C	C/C	A/A	A/A	A/A	C/C	A/A	A/A	A/A	A/A	A/A	C/C
c8	rs105694545	chr8	G/G	A/A	G/G	A/A	A/A	A/A	G/G	G/G	G/G	G/G	A/A	G/G	A/A	A/A
c9	rs65801920	chr9	T/T	T/T	C/C	C/C	T/T	C/C	T/T	T/T	C/C	T/T	T/T	T/T	T/T	T/T
c10	rs105168947	chr10	C/C	T/T	C/C	C/C	C/C	T/T	T/T	C/C	C/C	C/C	T/T	T/T	T/T	C/C
c11	rs106880606	chr11	G/G	T/T	T/T	G/G	T/T	T/T	T/T	G/G	T/T	T/T	T/T	T/T	T/T	G/G
d1	rs8158863	chr12	A/A	A/A	A/A	G/G	G/G	G/G	G/G	A/A	A/A	G/G	A/A	A/A	A/A	G/G
d2	rs105965886	chr13	C/C	T/T	T/T	T/T	T/T	C/C	C/C	C/C	T/T	C/C	C/C	C/C	C/C	C/C
d3	rs63919332	chr14	C/C	C/C	C/C	C/C	C/C	T/T	T/T	C/C	C/C	T/T	C/C	C/C	C/C	C/C
d4	rs65809306	chr15	T/T	T/T	T/T	T/T	T/T	C/C	C/C	T/T	T/T	C/C	T/T	T/T	T/T	C/C
d5	rs65087257	chr16	A/A^*	A/A	A/A	A/A	A/A	A/A	C/C	C/C^*	C/C	A/A	C/C	A/A	A/A	C/C
d6	rs106021355	chr17	T/T	C/C	T/T	T/T	T/T	C/C	T/T	T/T	T/T	T/T	T/T	T/T	T/T	T/T
d7	rs106171441	chr18	G/G	A/A	A/A	G/G	A/A	G/G	G/G	G/G	A/A	A/A	A/A	A/A	A/A	A/A
d8	rs106777713	chr19	G/G^*	A/A	A/A	G/G	A/A	T/T	C/C	T/T^*	T/T	C/C	C/C	T/T	T/T	T/T
d9	rs104937988	chr20	T/T	G/G	T/T	T/T	T/T	G/G	G/G	T/T	T/T	G/G	G/G	G/G	G/G	G/G
d10	rs105294203	chrX	G/G	G/G	A/A	G/G	G/G	A/A	G/G	G/G	A/A	G/G	G/G	G/G	G/G	G/G

基于多重PCR-LDR技术建立近交系大鼠单核苷酸多态性遗传检测方案

Establishing a Genetic Detection Protocol of Single Nucleotide Polymorphisms Panels in Inbred Rats Based on Multiplex PCR-LDR

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