16S amplicon sequencing of Fildes peninsula lake water
Surface water was collected using a 5-liter container. Each sample                 corresponds to 1-2 liters of water, filtered on a 0.22 m cellulose filter (Merck                 Millipore model GSQP04700) with a vacuum pump (Rocker 300). DNA was extracted from                 the filters using the DNeasy Power Water kit (Qiagen). The V4-V5 region of the 16S                 rRNA gene was amplified from 1:10 diluted DNA samples, using primers 515F and 928R                 (Wang & Qian, 2009) and Phusion Hot Start II High-Fidelity DNA Polymerase                 (Thermo Scientific), with annealing temperature of 72C. PCR products were paired-end                 sequenced on the Illumina Miseq platform (2 x 250-bp) (DNA Sequencing Facility,                 University of Wisconsin Biotechnology Center, Wisconsin, USA).