Extraction of purified plants gDNA free of Secondary metabolites

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Research Paper 01/03/2017
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Extraction of purified plants gDNA free of Secondary metabolites

Shahid Ali Khan, Niaz Ali, Gandeka Mamadou, Qinzheng Liu, Ruirong Zhuang, Hua Chen
Int. J. Biosci. 10(3), 223-231, March 2017.
Copyright Statement: Copyright 2017; The Author(s).
License: CC BY-NC 4.0

Abstract

Extraction of high-quality gDNA from plants is a basic step in molecular biology research. In some plant species like peanut getting the genomic DNA in its purified form is a very challenging job as compared to other crops. The reason behind which is the coexistence of different secondary metabolite impurities i.e., polyphenol and polysaccharides with genomic DNA. In this paper, we have optimized a protocol for extracting high-quality gDNA which is amenable to various improvement studies like PCR amplification and other molecular research. The OD 260/280 ratio for the extracted DNA was found within the range of 1.8 to 1.99 while OD 260/230 ratio was between 2.0 to 2.20, using Nano Drop ND-2000/2000C Spectrophotometer. Similarly, the concentration of the extracted DNA for all the extracted samples was greater than 300 ng/ul. The obtained results through our current method indicate the fitness of the extracted DNA for using it in PCR amplification and other molecular research. The novelty in our protocol is the use of Buffer A before CTAB treatment of the samples.

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