Download This PaperDesigning and Evaluation of Metagenomics 16S rRNA Gene Primers
14 Pages Posted: 6 Apr 2020 Last revised: 8 Jun 2020
Date Written: April 2, 2020
Abstract
Metagenomics is an emerging approach to study microbial diversity and its ecology to access nearly 100% of the environmental genomes. It helps to mine information about the genomes of both, cultivable and uncultivable microorganisms. Till date only limited microbial communities have been cultivated by traditional culturing methods. 16S rRNA gene is highly conserved and widely used to study the taxonomic status of the prokaryotes. In this study, salt enriched soil and seawater samples were collected from the coast of Kachhigadh, Dwarka and Alang, Bhavnagar. The metagenomic DNA was extracted by three different methods of soft lysis, harsh lysis, and combination of both. Combination of soft and harsh Lysis method yielded good quality of the metagenomic DNA with high purity. The extracted metagenomic DNA was intact with high molecular weight. Metagenomics primers for 16S rRNA gene were designed followed by the amplification of the concerned genes PCR products were then analyzed on the agarose gel electrophoresis. The DNA band size of the amplified products of 1500 and 700 bp were obtained by universal and metagenomic primers, respectively. The results suggested that the methods of the extraction of the metagenomic DNA were suitable for molecular downstream applications, such as PCR and cloning.
Keywords: Salt enriched soil, saline habitats, Seawater, Metagenomics DNA extraction, 16S rRNA gene, Metagenomics primers, PCR
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