DNA extraction from heavy oil contaminated microcosms and rpoB gene PCR amplification
DOI:
https://doi.org/10.17533/udea.acbi.329316Keywords:
DNA extraction, heavy oil, microcosms, PCR amplification, rpoBAbstract
This work describes a new method for extracting genomic DNA from heavy oil-contaminated soils. This method combines soil washes using three washing solutions with enzymatic lysis (SW-EL method). The suitability of the SW-EL method was validated by extracting DNA from samples as small as 0.5 g from model microcosms comprising soil artificially contaminated with 1 and 10% (w/w) heavy petroleum and seeded with a bacterial consortium. Compared with an established protocol for lysis and DNA purification using the commercial kit GeneReleaserTM (Bioventures) (GRL method), the novel SW-EL method provided higher DNA yields. With the addition of GeneReleaser to the PCR reaction to function as a chelating resin, rpoB genes (serving as model genes) could be successfully amplified. The SW-EL method has potential to be of use for DNA-based analysis of microbial consortia present in heavy-oil contaminated soils.
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