16S and 18S amplicon sequencing of Antarctic cryoconite holes
MoBio PowerSoil kits used to extract DNA from .3g (wet) sediments, and PowerWater kits used to extract DNA from Whatman nuclepore filters through which 50 ml melted water column was poured. Extracted genomic DNA was amplified in triplicate using 16S (515f-806r primers, Caporaso et al. 2012) and 18S (1391f-EukBr primers, Amaral-Zettler et al. 2009; Caporaso et al. 2012) SSU ribosomal gene markers. Amplified DNA was pooled and normalized to equimolar concentrations using SequalPrep Normalization Plate Kits (Invitrogen Corp., Carlsbad, CA, USA), and sequenced using the Illumina MiSeq V2 (2x250bp chemistry) at the BioFrontiers Sequencing Core Facility at the University of Colorado at Boulder.