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Volume: 2, Issue: 6, Nov-Dec, 2014
Amplification and cloning of mcrAgene from enriched consortia of Methanogens
Rachana Chandragupta Acharya, Usha Mukundan
Methanogens were enriched using anaerobic culture conditions. The gDNA extracted from the enriched consortia was used for amplification of mcrA (methyl coenzyme reductase A) gene. The temperature of annealing was standardized for amplification. The amplified gene was cloned into cloning vector and transformed into E.coli DH5α cells. Screening of recombinant transformants was done using Blue-White selection. The vector containing amplified gene was sequenced. The sequencing results showed that the gene had 97% homology with mcrA gene from Methanoculleus bourgensis. Thus procedure for mcrA gene amplification from enriched Methanogen consortia was standardized and its marker assisted identification using bioinformatics tools was carried out.
Keywords: Anaerobic culture, temperature of annealing, PCR amplification, marker assisted identification.