Asian Journal of Microbiology, Biotechnology & Environmental Sciences Paper

Vol.14, Issue 1, 2012; Page No.(129-136)

MOLECULAR ANALYSIS OF CRUDE OIL AND PRODUCED WATER FOR SULPHIDOGENIC BACTERIA

F.I. Akinnibosun and H.M. Lappin-Scott

Abstract

Molecular analysis of Nigerian Bonny light crude oil and Bonny light produced water (samples A and B respectively) was carried out to determine the presence of sulphide-generating bacteria (sulphidogenic bacteria). Samples were enriched in mixed carbon postgateĀ’s (MCP) medium and incubated at 550C for 28 days. DNA was extracted from enrichment and amplified by Polymerase Chain Reaction (PCR) using 16S rRNA Eubacterial primers with a GC-clamp. Successful amplification was confirmed by ethidium bromide fluorescence in 1% agarose gel during agarose gel electrophoresis. PCR-amplified DNA was separated by Denaturing Gradient Gel Electrophoresis (DGGE) analysis, to show the relationship between samples based on their GC content. Following successful DGGE analysis, DGGE bands were excised, purified and sequenced. Sequencing and phylogenetic analysis of DGGE products carried out to display sequence homology and affiliation to related genera of sulphide producers was successful displaying 98% and 99% similarity and homology to Petrotoga mexicana (AY125964.1) and Petrotoga olearia (AJ311703.1) respectively.

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