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Extraction of bulk DNA from Thar Desert soils for optimization of PCR-DGGE based microbial community analysis Electron. J. Biotechnol.
Kumar Gothwal,Raj; Kumar Nigam,Vinod; Mohan,M. Krishna; Sasmal,Dinakar; Ghosh,Purnendu.
A reliable method for characterizing microbial communities on the basis of their differences in the 16S ribosomal RNA (rRNA) gene sequences in the hot arid zone sandy soils has been optimized. A desert plant (Calligonum polygonoides) was chosen to provide the rhizospheric soil samples, collected from three different agro-ecological locations. Total community DNA was efficiently extracted at small-scale level using direct lysis with hot sodium dodecyl sulphate (SDS), glass bead beating and finally subjecting the sandy soil to liquid nitrogen freeze-thaw cycles. To amplify V3 region of bacterial 16S rRNA gene, universal conserved primers were used. Second round polymerase chain reaction (PCR) was attempted to increase product concentration and to minimize...
Tipo: Journal article Palavras-chave: Arid soils; Microbial community; PCR-DGGE; Rhizosphere; Soil DNA extraction.
Ano: 2007 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582007000300007
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