Johns M B, Paulus-Thomas J E
Department of Medical Genetics, Western Pennsylvania Hospital, Pittsburgh 15224.
Anal Biochem. 1989 Aug 1;180(2):276-8. doi: 10.1016/0003-2697(89)90430-2.
We have developed a new, rapid method for the extraction of human genomic DNA from whole blood samples. Traditionally, genomic DNA has been extracted from blood by overnight proteinase K digestion of lysed peripheral lymphocytes followed by phenol/chloroform extraction. In addition to being time consuming, the use of phenol involves inherent risks due to the toxic nature of the reagent. Our method for the extraction of DNA from whole blood uses sodium perchlorate and chloroform instead of phenol with a significant time savings realized as well as fewer hazards to the technician. Furthermore, DNA prepared by this new method is an excellent substrate for restriction endonuclease digestion and Southern hybridization analysis.
我们开发了一种全新的快速方法,用于从全血样本中提取人类基因组DNA。传统上,基因组DNA是通过对裂解的外周淋巴细胞进行蛋白酶K过夜消化,然后进行苯酚/氯仿提取从血液中提取的。除了耗时之外,由于该试剂的毒性,使用苯酚存在内在风险。我们从全血中提取DNA的方法使用高氯酸钠和氯仿代替苯酚,不仅显著节省了时间,而且对技术人员的危害也更小。此外,用这种新方法制备的DNA是限制性内切酶消化和Southern杂交分析的优质底物。