AN INTEGRATED STUDY OF A HOT SPRINGS MICROBIAL COMMUNITY IN THERMOPOLIS, WY
In preparation for amplification (PCR) and genetic sequencing the DNA was extracted using the PowerSoil DNA Isolation Kit from Mo Bio Laboratories, Inc. Once the DNA extraction process was completed the samples were placed into a -70oC freezer for storage until sent for off-site processing. The 16S rRNA genomes were sequenced by Research Testing Laboratory (RTL) of Lubbock, Texas. One-step PCR and Reads 454 MSC sequencing was performed utilizing Roche 454 FLX instruments and Titanium reagents to produce a tag-encoded FLX amplicon pyrosequencing analysis, and identified by comparisons with the NCBI (v.1-1-11) archives.
Results of sequencing show a microbial community populated by over 200 possible species, with moderate to high variability between sites. Dominant species include Chloroflexus sp and Spirulina sp with others identified as Pedobacter saltans, Thiofaba tepidiphila, Sulfurovum sp, Venenivibrio stagnispumantis, Enterobacter asburiae::Enterobacter sp, Flavobacterium sp, Sporobacterium sp. In addition to the microbial analysis, the distribution of microbe mats were mapped, water geochemistry was analyzed (and was consistent with earlier work) and underwater photography of the source spring was collected.