Development of a Membrane-Based Method for Isolation of Genomic DNA from Human Blood

Abstract

High quality and sufficient quantity of genomic DNA (gDNA) are the primary requisites of several molecular biologic applications, including clinical studies related to genetics, genomics, gene polymorphism, and DNA fingerprinting. Whole blood is the primary source of gDNA in most of the clinical investigations. Currently, commercial kits are primarily used to achieve these goals. However, the use of kits is limited by the cost and involvement of several centrifugal steps. Other methods reported are either laborious or do not produce high quality or quantity of gDNA or both. Here, we present the data on the development of a centrifugation-free, cost-effective, and user-friendly method for the isolation of human gDNA from the buffy coat of human blood that involves limited numbers of steps with about 15 min of hands-on time per sample.

Comparison of paper-based nucleic acid extraction materials for point-of-care testing applications

“…The PES membrane was prepared according to the reported literature (Mandal et al 2018 ) with slight modification. 30 μL of whole blood, 3 μL of protein K and 30 μL of lysis buffer (30 mM Tris-Cl, pH 8.0, 50 mM EDTA, 5% Tween 20, 0.5% Triton X-100, 1 M GuSCN) were added in EP tube.…”

“…Paper, as a low-cost, abundant, portable, biocompatible and easy-to-modify material, has been widely used for fabricating paper-based NAE device (Batule et al 2020 ; Mason and Botella 2020 ), which exhibits satisfactory performance for enabling nucleic acid detection at POC due to ease of use and cold chain-free transfer and storage. Common paper-based NAE devices are mainly based on various types of paper-based materials through nucleic acid specific adsorption and desorption, including Whatman filter paper #1 (Ghosh and Hazarika 2018 ; Hashimoto et al 2019 ; Zou et al 2017 ), FTA card (Choi et al 2016 ), FTA elute card (Hashimoto et al 2019 ), Fusion 5 (Jangam et al 2009 ), silica membrane (Tang et al 2019 ) and polyethersulfone (PES) membrane (Mandal et al 2018 ; Rodriguez et al 2015 ). For instance, Fusion 5-based automated DNA extraction device can extract DNA from whole blood within 8 min (Gan et al 2014 ).…”