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作 者:刘颖[1] 孙蓓[2] 马彦[2] 赵荣兰[2] 辛兢[2] 郭刚[2] 张镜宇[2]
机构地区:[1]天津医科大学口腔医院,300070 [2]天津医科大学内分泌研究所,卫生部及天津市激素与发育重点实验室
出 处:《天津医药》2008年第7期510-512,共3页Tianjin Medical Journal
摘 要: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.
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