STR等位基因梯阶制备的研究

A study on the preparation of STR allelic ladder

目的 建立制备STR的等位基因梯阶标准对照的新方法。方法 以T-质粒载体直接连接STR等位基因扩增产物,导入大肠杆菌,使其随大肠杆菌的无性繁殖而复制、扩增,获得单一DNA分子的大量拷贝;经PCR鉴定及测序分析后,制备出标准的STR等位基因梯阶对照(AL)。结果 所建方法制备出了标准AL;保存重组质粒及转染的大肠杆菌,可保证AL的大量快速制备,并能长期保存。结论 该方法制备的AL在法医物证检验中有很高的应用价值,对STR试剂盒国产化有重要意义。

Objective To produce the standard allelic ladder by using the cloning technique. Methods After the amplification and separation of the STR alleles, they were purified and then connected with T-vectors directly. The combinants were transfected into the component E. coli DH5α cells follwed by cloning and plasmid purification. The allelic ladder were then produced by re-amplifying the recombinant plasmid DNA. Results The allelic ladder made in this way can be produced in a lager amount and can be stored in a relatively long period. Conclusion The results demonstrated that the standard allelic ladder generated in this way is more practical in forensic scienc application. This technique in useful for preparation of domestic STR kits.