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International Journal of Current Microbiology and Applied Sciences (IJCMAS)
IJCMAS is now DOI (CrossRef) registered Research Journal. The DOIs are assigned to all published IJCMAS Articles.
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National Academy of Agricultural Sciences (NAAS)
NAAS Score: *5.38 (2020)
[Effective from January 1, 2020]
For more details click here

ICV 2018: 95.39
Index Copernicus ICI Journals Master List 2018 - IJCMAS--ICV 2018: 95.39
For more details click here

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Original Research Articles

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

Int.J.Curr.Microbiol.App.Sci.2017.6(9): 2695-2703
DOI: 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 Ambawat1,2, Ram Kumar1*, Subaran Singh1,2 and Rajesh Yadav1
1Department of Genetics and Plant Breeding, Chaudhary Charan Singh Haryana Agricultural University, Hisar 125004, Haryana, India
2Department of Agriculture Sciences and Research, SGV University,Jaipur-302 017, Rajasthan, India
*Corresponding author
Abstract:

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.
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How to cite this article:

Supriya Ambawat, Ram Kumar, Subaran Singh and Rajesh Yadav. 2017. An Easy, Quick and Cost Effective Method of High Quality DNA Extraction from Mungbean [Vigna radiata (L.) Wilczek] Without Liquid NitrogenInt.J.Curr.Microbiol.App.Sci. 6(9): 2695-2703. doi: https://doi.org/10.20546/ijcmas.2017.609.332