COI Metabarcoding of Arctic Periglacial Environments: moss; Svalbard
The COI gene was amplified from each of the 13 samples in duplicate, using the primers described in Folmer et al., 1994 and Dabert et al., 2015, with barcode on the forward primer at MR DNA company (www.mrdnalab.com, Shallowater, TX, USA). Thermocycling profile was set to one cycle of 5 min at 95 C followed by 35 steps of 30 s at 95 C, 90 s at 50 C, 1 min at 72 C, and a final step of 5 min at 72 C using the HotStarTaq Plus Master Mix Kit (Qiagen, USA). PCR products were checked in 2% agarose gel and samples were pooled together in equal proportions based on their molecular weight and DNA concentrations. Pooled samples were purified using calibrated Ampure XP beads and used to prepare Illumina DNA library. Libraries were sequenced on a MiSeq using 2x300 bp chemistry following the manufacturers guidelines.