Table of Contents
Molecular Biology International
Volume 2014 (2014), Article ID 518960, 6 pages
http://dx.doi.org/10.1155/2014/518960
Research Article

An Improved Method for Soil DNA Extraction to Study the Microbial Assortment within Rhizospheric Region

1Department of Biosciences, Integral University, Lucknow 226026, India
2Department of Biotechnology, Delhi Technological University, Delhi 110042, India

Received 18 May 2014; Revised 10 August 2014; Accepted 19 August 2014; Published 15 September 2014

Academic Editor: Malayannan B. Subramaniam

Copyright © 2014 Faria Fatima et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

The need for identification of soil microbial community mainly depends on direct extraction of DNA from soil, a multifaceted environment that is a major pool for microbial genetic diversity. The soil DNA extraction procedures usually suffer from two major problems, namely, inappropriate rupturing of cells and contamination with humic substances. In the present study, five protocols for single type of rhizospheric soil were investigated and their comparison indicated that the inclusion of 120 mM phosphate buffered saline (PBS) for washing and mannitol in the lysis buffer allowed the processing of soil sample in minimal time with no specific equipment requirement. Furthermore, DNA purity and yield were also improved, which allowed the exploitation of genetic potential of soil microbes within soil sample thereby facilitating the amplification of metagenomic DNA. The effectiveness of methods was analyzed using random amplification of polymorphic DNA. The banding patterns revealed that both the abundance and the composition of indigenous microbial community depend on the DNA recovery method.