16S metagenomics: Slovenia, Paradana cave, cave ice-1
Total community DNA was isolated using MoBio PowerWater DNA Isolation kit. The 16S region of total community DNA was amplified with 16S Ion Metagenomics Kit. DNA library was made using Ion Plus Fragment Library Kit and Ion Xpress Barcodes Adapters. Size distribution and number of processed libraries were evaluated with the Labchip GX instrument (Perkin Elmer). The concentration of each sample was adjusted to 20 pM. Equal volumes of all samples were combined and processed in emulsion PCR and enrichment steps with One-Touch 2 and One-Touch ES systems using the Ion PGM Hi-Q View OT2 Kit. The libraries were sequenced in a 316 v2 chip on the Ion Personal Genome Machine using the Ion PGM Hi-Q View Sequencing Kit.