Loparev V N, Cartas M A, Monken C E, Velpandi A, Srinivasan A
Wistar Institute of Anatomy and Biology, Philadelphia, Pennsylvania 19104.
J Virol Methods. 1991 Sep;34(1):105-12. doi: 10.1016/0166-0934(91)90126-k.
Routine methods of extraction of DNA from blood involve the enrichment of cells by Ficoll-Hypaque gradient centrifugation followed by lysis of the cells with extraction buffer, proteinase K digestion of the lysate, and phenol:chloroform-isoamyl alcohol extraction. These methods generally require large amounts of blood, which poses a problem with pediatric patients. To overcome this, we developed a new method of extracting DNA directly from whole blood. This method involves the treatment of whole blood with an equal volume of NaI (3 M final concentration) followed by chloroform:isoamyl alcohol extraction to clear hemoglobin and cell debris. The clear aqueous layer is then mixed with isopropanol to obtain DNA. A large number of samples can easily be handled by this extraction procedure, as it can be carried out in 30 min and requires only a microcentrifuge.
从血液中提取DNA的常规方法包括通过Ficoll-Hypaque梯度离心富集细胞，随后用提取缓冲液裂解细胞，对裂解物进行蛋白酶K消化，以及苯酚：氯仿-异戊醇提取。这些方法通常需要大量血液，这对儿科患者来说是个问题。为克服这一问题，我们开发了一种直接从全血中提取DNA的新方法。该方法包括用等体积的NaI（终浓度3M）处理全血，随后进行氯仿：异戊醇提取以清除血红蛋白和细胞碎片。然后将清亮的水相层与异丙醇混合以获得DNA。这一提取程序能轻松处理大量样本，因为它可在30分钟内完成，且仅需一台微量离心机。
