Int.J.Curr.Microbiol.App.Sci (2017) 6(9): 2695-2703 2695 Original Research Article https://doi.org/10.20546/ijcmas.2017.609.332 An Easy, Quick and Cost Effective Method of High Quality DNA Extraction from Mungbean [Vigna radiata (L.) Wilczek] Without Liquid Nitrogen Supriya Ambawat 1,2 , Ram Kumar 1* , Subaran Singh 1,2 and Rajesh Yadav 1 1 Department of Genetics and Plant Breeding, Chaudhary Charan Singh Haryana Agricultural University, Hisar 125004, Haryana, India 2 Department of Agriculture Sciences and Research, SGV University, Jaipur-302 017, Rajasthan, India *Corresponding author ABSTRACT Introduction Green gram (Vigna radiata (L.) Wilczek) or mungbean is a self-pollinated widely cultivated leguminous crop in India. It belongs to the family Leguminaceae and subgenus Ceratotropis with diploid chromosome number (2n=2x=22) (Kang et al., 2014). It is mainly grown in tropical Africa and Asia and several Vigna species have been domesticated in Asia. It is an important source of protein for the human population and soil health as it fixes atmospheric nitrogen. High protein, easy digestibility and low flatulence make it widely acceptable to the people world over. The annual world production area of mungbean is about 5.5 million hectare of which about 90% is in Asia (Lambrides and Godwin, 2007). India is the largest producer of mungbean and occupies an area of 34.4 lakh hectares with a production of 1.60 million tonnes and productivity of 568 kg/ha (2015-2016). Genus Vigna have high amount of polyphenol, orthohydroxyphenols and polysaccharides. International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 6 Number 9 (2017) pp. 2695-2703 Journal homepage: http://www.ijcmas.com Green gram is a widely cultivated pulse crop rich in protein, essential amino acids and vitamin-B. The quality and quantity of DNA are very important for amplification by PCR. Although quantity of DNA required per reaction in PCR is very low, quality is very crucial. Also, to carry out large number of PCR reactions for genotyping, a good amount of DNA is required. Presence of contaminants like phenols makes it difficult to get good quality DNA from mungbean. Thus, the present study was undertaken to obtain high quality and pure DNA in mungbean. The method involves extraction of DNA using a buffer (pH 8.0) containing 100mM Tris, 50mM EDTA, 500mM NaCl, 2%PVP, 2%CTAB and 0.2%β-mercaptoethanol followed by purification of DNA with chloroform and isoamyl alcohol and finally precipitation of DNA by sodium acetate and isopropanol. The method is suitable for extraction of DNA from small to large number of plant samples. DNA obtained through this protocol is of high quality and free of phenols which gave amplifying products in the PCR. Here, we developed a simple, fast, efficient, economical method for isolation of DNA from green gram without liquid nitrogen which could be stored for longer duration and does not require expensive chemicals such as proteinase K, liquid nitrogen etc. Keywords Green gram, DNA extraction, Standardization, Phenols, Liquid nitrogen. Accepted: 26 August 2017 Available Online: 26 September 2017 Article Info