A rapid and efficient DNA extraction protocol from fresh and frozen human blood samples

作者：Guha Pokhraj; Das Avishek; Dutta Somit; Chaudhuri Tapas Kumar^*

来源：Journal of Clinical Laboratory Analysis, 2018, 32(1): e22181.

DOI：10.1002/jcla.22181

• cost effective
• DNA extraction
• high yield
• human blood

BackgroundDifferent methods available for extraction of human genomic DNA suffer from one or more drawbacks including low yield, compromised quality, cost, time consumption, use of toxic organic solvents, and many more. Herein, we aimed to develop a method to extract DNA from 500L of fresh or frozen human blood.
MethodsFive hundred microliters of fresh and frozen human blood samples were used for standardization of the extraction procedure. Absorbance at 260 and 280nm, respectively, (A(260)/A(280)) were estimated to check the quality and quantity of the extracted DNA sample. Qualitative assessment of the extracted DNA was checked by Polymerase Chain reaction and double digestion of the DNA sample.
ResultsOur protocol resulted in average yield of 222.97g and 20.5 +/- 3.97g from 500L of fresh and frozen blood, respectively, which were comparable to many reference protocols and kits.
ConclusionBesides yielding bulk amount of DNA, our protocol is rapid, economical, and avoids toxic organic solvents such as Phenol. Due to unaffected quality, the DNA is suitable for downstream applications. The protocol may also be useful for pursuing basic molecular researches in laboratories having limited funds.

• 出版日期2018-1