18S amplicon sequencing of Dinwoody Glacier sediment: Din24
DNA was extracted from soils using the DNeasy PowerSoil Kit (Qiagen, Hilden, Germany) and DNA concentrations were assayed with a Qubit fluorometer (Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturers directions. Marker genes were amplified by the Fierer lab at the University of Colorado Boulder using the Earth Microbiome Project primers 515f-806r (Caporaso et al. 2011, Parada et al. 2016) and 1391-EukBr (Amaral-Zettler et al. 2009) to detect bacteria and eukaryotes respectively, and normalized to equimolar concentrations and the addition of barcodes. Libraries were then sequenced on an Illumina MiSeq at the Biofrontiers Sequencing Facility at the University of Colorado Boulder at 2 x 250 bp chemistry for the 16S library and 2 x 150 bp chemistry for the 18S library.