18S amplicon sequencing McMurdo Dry Valleys and McMurdo Sound
RNA was extracted from frozen, RNAlater preserved samples using the protocol from the NucleoMag RNA (Macherey-Nagel) kits. 1-10 ng of total RNA was used to generate cDNA using the Life Technologies. SuperScript III First Strand Synthesis system with random hexamer primers. PCR with V4 primer set 1389 F (5-CCAGCASCYGCGGTAATTCC-3) and 1510 R (5-ACTTTCGTTCTTGATYRA-3); one-step PCR using the TruFi DNA Polymerase; purified using AMPure XP beads; quantified and pooled.