构建基于荧光多重 cPCR 的小鼠基因拷贝数检测方法

摘要

目的：建立基于竞争性聚合酶链式反应（ competitive polymerase chain reaction， cPCR）小鼠基因拷贝数变异（ copy number variations， CNVs）的检测方法，用于检测野生小家鼠来源一号染色体替换系群体（ population of specific chromosome 1 substitution strains， PCSSs）的CNVs。方法选取小鼠一号染色体上11个CNVs位点，及7、9和X染色体上各1个内对照位点，分别构建克隆质粒为竞争性粒模板，应用cPCR技术，建立荧光通用引物多重cPCR检测方法。结果多重cPCR方案适用于小鼠一号染色体上11个CNV位点的拷贝数检测，且能准确检测X染色体的拷贝数。结论实现小鼠快速、高通量的CNVs检测，可准确检测小鼠1号染色体中11个CNV位点的拷贝数变异。%Objective To establish a high throughput general multiple competitive polymerase chain reaction ( cPCR) detecting method of copy number variations ( CNVs) for the population of chromosome 1 substitution strains from wild mice.Method The selected 14 loci, including 11 CNVs on chromosome 1 and internal control loci on other three chromosmes (Chr 7, Chr 19 and Chr X), were detected based on the universal fluorescent primer multiple competitive pol-ymerase chain reaction.All specific cloned plasmids were constructed as competitors.Results Altogether 11 CNVs were designed in one panel, and the copy of Chr X accurately reflects the gender.Conclusions A rapid and high-throughput fluorescent multiplex cPCR assay is established which can be used for detection of copy number variations on chromosome 1 in mice.