Research Article | Published: 01 September 2015

Genomic DNA isolation from old needles of Cedrus deodara (Roxb.) G.Don and amplification of nuclear microsatellite markers

Akhilesh  Kumar, Santan Barthwal and Harish Singh Ginwal

Indian Journal of Forestry | Volume: 38 | Issue: 3 | Page No. 215-217 | 2015
DOI: https://doi.org/10.54207/bsmps1000-2015-8371AD | Cite this article

Abstract

A reliable and modified protocol based on the CTAB method was developed for DNA extraction from old needles of Cedrus deodara (Roxb.) G.Don. The presence of high concentrations of polysaccharides, polyphenols and other secondary metabolites in C. deodara needles poses problem in getting good quality DNA fit for PCR applications. used 2.5% PVP for removal of phenolics compound and apply high concentration of sodium chloride to removes polysaccharides. Oils was removed by centrifugation. The yield of extracted DNA was ranged from 35 to 130 µg/100mg fresh weight of the needles with absorbance ratio (A260/A280) ranged from 1.82 to 2.0. DNA extracted by modified protocol gives positive amplification with PCR using Nuclear Simple Sequence Repeat (SSR) markers. This modified protocol can be used in genomic DNA extraction from closely related species of C. deodara.

Keywords

CTAB, Nuclear Simple Sequence Repeat (SSR), DNA extraction, PCR application, Cedrus deodara (Roxb.) G.Don Needles

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How to cite

Kumar, A., Barthwal, S. and Ginwal, H.S., 2015. Genomic DNA isolation from old needles of Cedrus deodara (Roxb.) G.Don and amplification of nuclear microsatellite markers. Indian Journal of Forestry, 38(3), pp.215-217. https://doi.org/10.54207/bsmps1000-2015-8371AD

Publication History

Manuscript Published on 01 September 2015

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