从一根毛发中分别提取nDNA和mtDNA的简易方法

The Study of the DNA Extraction Procedure from a Single Hair

目的:探讨从1根脱落毛发中既获取核内基因组DNA(nDNA)又获取核外线粒体DNA(mtDNA)的简便可靠方法。方法:用含高浓度的二硫苏糖醇(DTT)的细胞裂解液裂解细胞,建立DTT-裂解液法及DTT-TKM法用于毛根、毛干部分的核酸提取,并对2种方法进行了比较。结果:2种方法均能从毛根及毛干中提取nDNA及mtDNA。DTT-TKM法适用于多根毛发的核酸提取。而DTT-裂解液法则不仅能提高毛囊DNA的提取量,而且能从少量毛干中提取到几乎不含nDNA的mtDNA。结论:DTT-裂解液法简便、经济、无创,更适于从1根毛发中分别提取nDNA及mtDNA。特别适用于血液等样本采集困难时对于遗传信息的获取,在法医学上亦具有一定的应用价值。

Objective： To establish a reliable and brief procedure for extracting the nuclear DNA （nDNA） and mitochondrial DNA （mtDNA） from a single shed hair respectively. Methods： Two new methods were established and compared with DTT addition to the cell lysis buffer for DNA extracting. Nucleic acid was analyzed by PCR and electrophoresis. Results： Both two procedures are able to extract nDNA and mtDNA from the hair follicle and shaft. Method of DTT-TKM is suitable for more hair samples,while method of DTT-lysis buffer can improve the quantity of the DNA extraction from a single hair follicle and can also extract mtDNA with scarcely any nDNA from a single hair shaft. Conclusion： The procedure with DTT-lysis buffer is more suitable for extracting nucleic acid from a single shed hair.