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Please use this identifier to cite or link to this item: http://hdl.handle.net/123456789/9913

Title: Extraction of Microbial Community DNA from Soil for Polymerase Chain Reaction
Authors: Wafaa M. H. Zidan
M. A. Radwan
H. El-Khawas
M. K. Zahra
S. M. Badr El-Din
Issue Date: 2005
Publisher: Egyptian Journal of Applied Science
Abstract: The development of techniques in molecular biology has led to their application to microbial ecology. The extraction of DNA, followed by the use of Polymerase Chain Reaction (PCR) to amplify a gene common to all organisms can provide information about microbial community structure, microbial diversity, evolution and taxonomy. The gene commonly amplified for this purpose codes for the RNA sequence of the small subunit (SSU) of the ribosome. Two methods, namely SDS-based extraction method and FastPrepTM method, were evaluated using four different soil samples represent major types of Egyptian soils. The DNA fragment size in the extracts from all soils was >23 kb. The DNA yield from the 4 soils ranged from 18 -52 µg of DNA g-1. The highest DNA yield was observed in Beni-Suef soil rich in organic carbon and nitrogen content and high microbial population. However, Ismalia soil, which is low in organic carbon, nitrogen content and microbial population, had the lowest DNA yield. DNA extracts by SDS based method from all soils did not exhibit high absorbance ratios, due to the presence of absorbing contaminants, primarily humic acids which had inhibited subsequent molecular reactions such as PCR amplification. FastPrepTM method, based on a mechanical lysis with ceramic and silica beads in a bead-beater, resulted in very pure, high molecular weight, and unsheared DNA suitable for PCR amplification.
URI: http://hdl.handle.net/123456789/9913
Appears in Collections:College of Foods And Agricultural Science

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