National Academy of Agricultural Sciences (NAAS)
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PRINT ISSN : 2319-7692
Online ISSN : 2319-7706 Issues : 12 per year Publisher : Excellent Publishers Email : editorijcmas@gmail.com / submit@ijcmas.com Editor-in-chief: Dr.M.Prakash Index Copernicus ICV 2018: 95.39 NAAS RATING 2020: 5.38 |
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.
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