等位基因测序的简便快速方法

目的]探索一种简便快速的等位基因测序方法.方法]用银染方法对待检个体的血样本进行简短串联重复(shorttandemrepeat,STR)基因分型,将需测序的等位基因条带从凝胶图上切下,PCR扩增,产物经纯化作为测序模板.采用循环测序法测序.结果]对D12S391、D11S554STR基因座的等位基因Ladder及STR基因座(FGA、D12S391、D11S554)等位基因发生突变的19个家系的等位基因进行了测序.STR分型表现为杂合子或纯合子的等位基因用此方法测序都得到了清晰的信号.结论]此方法不仅给等位基因测序提供了一种有效手段,而且对于只要能利用电泳分离的混合的DNA片段,就能用此方法将各DNA片段分别测序.

One Simple and Rapid Method for Allele Sequencing

To develope a simple and rapid method for allele sequencing. The blood samples of individuals were genotyped by silver staining on short tandem repeats(STR). Alleles to be sequenced were excised from gels, reamplified by PCR, and purified. Sequencing was performed by cycle sequencing method. Good signals had been obtained, no matter the alleles showed homozygotes or heterozygotes on STR genotyping. Conclusion] This method provides a good tool for allele sequencing and so long as the mixed DNA fragments can be separated by electrophoresis, the DNA fragments could be sequenced respectively by this method.