title: 16S amplicon sequencing McMurdo Dry Valleys and McMurdo Sound
design: 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-V5 primer set 515 F (5-GTGYCAGCMGCCGCGGTAA-3) and 926 R (5-CCGYCAATTYMTTTRAGTTT-3); one-step PCR using the TruFi DNA Polymerase; purified using AMPure XP beads; quantified and pooled.
library_name: P1Miers_R_G_12_20_16_16S