An Easy, Quick and Cost Effective Method of High Quality DNA Extraction from Mungbean [Vigna radiata (L.) Wilczek] Without Liquid Nitrogen

Development of a High Quality, Rapid, Efficient and Economical DNA Extraction Protocol from Climate Resilient Pearl Millet Crop Without Liquid Nitrogen

“…Antioxidants like β mercaptoethanol, PVP, bovine serum albumin and sodium azide are mainly used to get rid of the problems associated with phenolics [17,32]. In the present study, the extracted DNA was free of phenols as the pellet obtained was clean, clear and translucent which is comparable to other such recent studies [2,18,19,36,37]. β mercaptoethanol acts as a reducing agent and removes the tannins and polypheno crude extract by denaturing proteins and During extraction process, cell wall is broken down first of all followed by digestion and disruption of cell membrane to release the cellular constituents and DNA into the extraction buffer using detergents like sodium dodecyl methylammonium Here, we used combination of both detergents, CTAB and SDS to make extraction more efficient in absence of liquid nitrogen.…”

“…Antioxidants like βmercaptoethanol, PVP, bovine serum albumin and sodium azide are mainly used to get rid of the problems associated with phenolics [17,32]. present study, the extracted DNA was free of phenols as the pellet obtained was clean, clear and translucent which is comparable to other such recent studies [2,18,19,36,37]. βa reducing agent and removes the tannins and polyphenols from the crude extract by denaturing proteins and breaking the disulfide bonds between the cysteine residues.…”

“…In addition, on analysis on agarose gel electrophoresis, sharp and distinct bands were observed with no visible DNA degradation or RNA contamination indicating good quality and quantity of extracted genomic DNA (Fig.1). The good quality of DNA extracted in our study is comparable to various other studies where it was reported that good quality DNA can be isolated without using liquid nitrogen [19,21,22,24]. Most DNA extraction methods are time consuming, involve multiple steps, expensive chemicals and enzymes and use hazardous procedures of grinding plant tissue in liquid nitrogen (N 2 ) to break down the cell wall of plants [21] or freeze-drying (lyophilization) [25].…”

“…The yield of extracted DNA ranged from 105.2 to 328.3 ng/μl and the ratio of A 260/280 varied from1.74 to 1.95 specifying good quality DNA which is free from protein and RNA contamination (Table 2). Similar kind of yield and quality of DNA was also reported in other protocols [2,4,19,21,22]. A ratio of absorbance of 260/280 is used to judge the purity of nucleic acid and DNA quality with an OD 260/ OD 280 ratio between 1.7 and 2.0 is supposed to be good [23].…”

“…The isolated DNA was analyzed on 0.8% agarose gel electrophoresis by loading a mixture of 1μl DNA sample and 4 μl of 6X loading dye along with 50 bp ladder [19]. The gel was run at 80V, till the tracking dye migrated to the bottom of the gel.…”