rRNA gene of eukaryotes: Antarctic cryoconite hole water column
DNA was extracted with MoBio PowerWater DNA Extraction Kit according to directions. Extracted genomic DNA was amplified in triplicate using 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, United States), and sequenced using 2  250 bp chemistry) at the BioFrontiers Sequencing CoreFacility at the University of Colorado at Boulder.