title: Bacteria community analysis using Nanopore
design: The DNA extraction was performed as described above. Bacterial community analyses of Antarctic soil samples were performed using Oxford NanoporeTM sequencing (Oxford Nanopore Technologies, U.K.). PCR reactions were performed using 10 ng of DNA from each sample, in addition to 25 L of Long Amp Hot Start Taq 2X Master Mix and a 16S-specific barcoded primer pair (27F; AGA GTT TGA TCC TGG CTC AG, 1492R; GGT TAC CTT GTT ACG ACT T). The DNA samples were then purified using a 16S barcoding kit (SQK-16S024). DNA (100 ng/L) was obtained by pooling equal concentrations of DNA for each sample, after this sequencing was performed. The sequencing procedures rendered read counts of approximately Gigabases (Gb) using a single flow cell on a MinION portable sequencer (Oxford Nanopore Technologies, U.K.). Adapter trimming and assembly of raw sequences were performed using the CLC Genomic Workbench software version 20 (Qiagen, Germany).