反向PCR:缺失断点定位好方法

The inverse PCR:good method for deletion breakpoint mapping

目的利用反向聚合酶链式反应(PCR)对乳腺癌细胞系MCF-7的9p21缺失断点进行准确定位。方法以乳腺癌MCF-7细胞系为模板,利用反向PCR,通过酶切、连接、PCR反应及测序,检测染色体9p21缺失断点。结果乳腺癌MCF-7细胞系9p21缺失断点起于chr9:21819532、止于chr9:21989622的大小为170kb的大片段缺失,断点处可见ACTGG 5个碱基的插入,与长片段PCR检查结果相符。结论反向PCR是缺失断点定位好方法,适合样本人群研究。

Objective To map the deletion breakpoint of 9p21in breast cell line MCF-7use the inverse PCR.Methods After digestion,ligation and PCR reaction,the breakpoint was confirmed by sequencing.Results The deletion breakpoint started at chr9： 21819532and ended at chr9：21989622with a small insertion of ACTGG,which was consistent with the result confirmed by the long range PCR.Conclusion The inverse PCR is one good method for deletion breakpoint mapping and suitable for large sample size detection.