Establishing a Genetic Detection Protocol of Single Nucleotide Polymorphisms Panels in Inbred Rats Based on Multiplex PCR-LDR
10.12300/j.issn.1674-5817.2023.073
- VernacularTitle: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
- Author:
Liya ZHAO
1
;
Liju NI
1
;
Caiqin ZHANG
2
;
Jianping TANG
1
;
Yangzheng YAO
3
;
Yanyan NIE
1
;
Xiaoxue GU
4
;
Ying ZHAO
1
Author Information
1. Shanghai Laboratory Animal Research Center, Shanghai 201203, China
2. Laboratory Animal Center of Air Force Medical University, Xi'an 710038, China
3. Shaanxi Academy of Traditional Chinese Medicine, Xi'an 710003, China
4. Shanghai BK/KY Biotechnology Co., Ltd.,Shanghai 201203, China
- Publication Type:Journal Article
- Keywords:
Inbred rats;
Single Nucleotide Polymorphisms;
Multiplex PCR-LDR;
Genetic detection;
Verification
- From:
Laboratory Animal and Comparative Medicine
2023;43(5):548-558
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo establish a set of single nucleotide polymorphisms (SNP) detection protocol for inbred rats based on multiplex PCR-ligase detection reaction (LDR). MethodsA 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. ResultsWhen 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. ConclusionBased 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.
