基于PCR-LDR平台的近交系小鼠遗传质量快速检测方法

目的建立基于PCR-LDR平台的近交系小鼠SNP快速分型方法,用于检测实验小鼠的遗传质量与品系纯度。方法利用可移植性极高的PCR-LDR技术,以常见近交系小鼠为研究对象,选取了21条染色体上的45个SNP位点,分别设计引物和探针,经过筛选和验证,建立了多重PCR-LDR(polymerase chain reaction and ligase detection reaction,PCR-LDR)分型方案。结果四组多重PCR-LDR可实现45个SNP位点的基因分型,其中43个、44个与45个SNP在样本中的检出率分别为100%、90.9%与36.4%。所有样本经分型确定为纯合体,并得到了常见近交系小鼠SNP位点信息。结论实现了常见近交系小鼠快速、高通量的基因分型,可用于遗传质量检测和品系鉴定。

A rapid detection method of inbred mouse DNA genetic quality based on PCR-LDR platform

Objrctive Purity of laboratory animals plays a critical role affecting the experimental results.The aim of this study was to validate a high-throughput genotyping platform,valuable in inbred mouse genetic DNA monitoring and strain identification.Methods Multiple polymerase chain reaction and ligase detection reaction(PCR-LDR) genotyping panels were constructed.Forty-five single-nucleotide polymorphism(SNP) on 21 chromosomes were selected as targets,as well as specific primers and probes to these SNP were designed,respectively.A multiple PCR-LDR genotyping protocol was established.Results Forty-five SNP were successfully genotyped in four panels.The positive detection rate of 43,44 and 45 SNPs were 100%,90.9% and 36.4%,respectively.All samples collected were homozygous and their genotypes were identified by PCR-LDR.Conclusion The results of this study provide a rapid and high-throughput genotyping approach,which is sufficient for genetic contamination monitoring and strain identification.