从极少量人血凝块中快速提取基因组DNA的方法

Fast method of extracting DNA from very few human blood clots

目的建立从极少量血凝块中快速提取基因组DNA的方法,并对其应用价值进行评价。方法分别对TIANamp基因组DNA提取试剂盒和TaKaRa全能基因组DNA提取试剂盒进行方法改良,并提取DNA,比较2种方法提取DNA的含量和纯度;并用人P450酶系的CYP3A4*4引物对提取的DNA进行PCR扩增,检测生物活性。结果用TaKaRa全能基因组DNA提取试剂盒提取的DNA在含量、纯度和生物活性上均优于用TIANamp基因组DNA提取试剂盒提取的DNA。结论应用TaKaRa全能基因组DNA提取试剂盒可以对极少量的血凝块进行基因组DNA提取,方法简便快速,在分子生物学研究中有重要的实际意义和推广价值。

AIM To establish a fast method of extracting DNA from very few blood clots, and evaluate its application value. METHODS DNA was extracted respectively by using TIAINamp Genomic DNA Kit and TaKaRa Universal Genomic DNA Extraction Kit which both were improved. The content and purity of DNA were compared. The biological activity was detected by using PCR with CYP3A4 ~ 4 primers. RESULTS The content, purity and biological activity of DNA that was extracted with TaKaRa Universal Genomic DNA Extraction Kit were all better than the DNA extracted with TIANamp Genomic DNA Kit. CONCLUSION It is practical to extract DNA from very few blood clots with TaKaRa Universal Genomic DNA Extraction Kit. The method is simple and fast, and has an important significance in research of molecular biology.