Dataset: Microbial community composition McMurdo Spiophanes beds
Deployment: Thurber_Hutt_Point_2012

Methodology:

Sediment cores were collected from the Spiophanes beds at the “Jetty” dive location and vertically sectioned into cm intervals with the exterior of the cores discarded to avoid vertical smearing.  Sediment were placed in whirlpack bags and kept at -80 until DNA was extracted. 
 
Sampling and analytical procedures:

 Between 0.25 and 0.5 grams of frozen sediment had DNA extracted using the MoBio (now Qiagen) PowerSoil kits. rimers and amplification follows the Earth Microbiome Project Protocol (http://www.earthmicrobiome.org/protocols-and-standards/16s/) using the updated primers in Apprill et al (2015) following Caporaso et al. (2011).  In short, triplicate PCRs were run using the 515FB and the 806RB primers that copy the V4 region if the 16s rRNA gene.  These primers were barcoded allowing later in silico separation of pooled samples.  Controls and all samples were run on a gel to check for contamination.  DNA was cleaned up using the MoBio UltraClean PCR Clean-Up Kit, samples were pooled into equal molar concentrations and submitted for sequencing. 

Sequencing was performed on the Illumina MiSeq platform using the V.2 chemistry at the Center for Genome Research and Biocomputing at Oregon State University.