IMIS

Soil samples were collected aseptically at each site into sterile Whirl-Pak bags using a sterile metal spatula and kept frozen during transport to the laboratory.
Study Extent: Samples were collected from the Miers Valley region of Antarctica during the summer months, including: MS1, Miers Stream 1 (S78°05.748′, E163°44.822′), a site located at the edge of the runoff stream flowing from Miers Glacier into Miers Lake in Miers Valley; ML1, Miers Lake 1 (S78°05.615′, E163°49.912′) located on the north side of Miers Lake; BL, Baby Buddha Lake (S78°03.621′, E163°46.463′), and NP, Nostoc Pond (S78°03.920′, E163°46.497′) adjacent to lake systems located over the north ridge of Miers Valley.
Method step description:

1. DNA was isolated from homogenized soil samples using the PowersoilTM DNA isolation kit (MOBIO) as per manufacturer’s instructions with all terminal-restriction fragment length polymorphisms (T-RFLPs) based on pooled triplicate PCR assays.
2. Tagged amplicon pyrosequencing of the V1 to V3 region of the 16S and 18S rRNA gene (primer pairs 28F-519R and euk7F-euk570R, respectively) was performed on DNA extracts by the Research and Testing Laboratories located in Lubbock, TX (www.researchandtesting.com) using FLX technology (Roche).