PURIFICATION OF HUMAN GENOMIC DNA FROM WHOLE-BLOOD USING SODIUM-PERCHLORATE IN PLACE OF PHENOL
PURIFICATION OF HUMAN GENOMIC DNA FROM WHOLE-BLOOD USING SODIUM-PERCHLORATE IN PLACE OF PHENOL
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DOI：
10.1016/0003-2697(89)90430-2
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发表时间：
1989-08-01
影响因子：
2.9
通讯作者：
PAULUSTHOMAS, JE
中科院分区：
文献类型：
--
作者：
JOHNS, MB;PAULUSTHOMAS, JE
我们开发了一种从全血样本中提取人类基因组DNA的新的快速方法。传统上，基因组DNA是通过蛋白酶K过夜消化裂解的外周淋巴细胞，然后用苯酚/氯仿提取而从血液中提取的。除了耗时之外，由于试剂的毒性性质，使用苯酚还涉及固有风险。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，节省了大量时间，对技术人员的危害也更小。此外，这种新方法制备的DNA是一个很好的限制性内切酶消化和Southern杂交分析的底物。
We have developed a new, rapid method for the extraction of human genomic DNA from whole blood samples. Traditionally, genomic DNA has been extractedfrom blood by overnight proteinase K digestion of lysed peripheral lymphocytes followed by phenol/chloro form 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.