A rapid and efficient DNA extraction protocol from fresh and frozen human blood samples. Issue 1 (23rd February 2017)

• Record Type: Journal Article
• Authors: Guha, Pokhraj; Das, Avishek; Dutta, Somit; Chaudhuri, Tapas Kumar
• Is Part Of: Journal of clinical laboratory analysis. Volume 32:Issue 1(2018)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2017-02-23
• Subjects: cost effective -- DNA extraction -- high yield -- human blood; Diagnosis, Laboratory -- Periodicals; Medical laboratory technology -- Periodicals; 616
• DOI: 10.1002/jcla.22181
• Languages: English
• ISSNs: 0887-8013

Abstract

Abstract : Background: Different 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 500 μL of fresh or frozen human blood. Methods: Five hundred microliters of fresh and frozen human blood samples were used for standardization of the extraction procedure. Absorbance at 260 and 280 nm, respectively, (A260 /A280 ) 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. Results: Our protocol resulted in average yield of 22±2.97 μg and 20.5±3.97 μg from 500 μL of fresh and frozen blood, respectively, which were comparable to many reference protocols and kits. Conclusion: Besides 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.