Microbial diversity of oil spill sands
"Total nucleic acids were extracted in triplicate. Five grams sediment for each sample was subjected to methods modified from Somerville et al. (1989; Appl Environ Microbiol, 55(3): 548-554) and Zhou et al. (1996, Appl Environ Microbiol, 62(2): 316-322) that incorporated sucrose lysis buffer with lysozyme (1 mg/ml), a solution of 5X proteinase K/CTAB/SDS, and overnight incubation at 55 C while shaking at 100 rpm. After protein precipitation, nucleic acids were precipitated in isopropanol overnight at -20?C prior to washing with ethanol and purification. Pyrosequencing of the V1-V3 region of 16S rRNA genes was done using a 454 GS FLX Titanium at the Research and Testing Laboratory (Lubbock, TX)."