Metatranscriptome sequencing Antarctica: McMurdo Dry Valleys
Total RNA was extracted from frozen, RNAlater preserved sample using the NucleoBond RNA Soil kit (Macherey-Nagel, Dren, Germany); cleanup and DNase treatment by RNA Clean & Concentrator (Zymo Research, Irvine, CA, USA).  Then, total RNA was subjected to amplification and cDNA synthesis using the Ovation RNA-Seq System V2 (TECAN, Redwood City, CA, USA). Double stranded cDNA was fragmented using Covaris E210 system with the target size of 300bp. Libraries were constructed by Ovation Ultralow System V2 (TECAN) following the manufacturer's protocol. Ampure XP beads (Beckman Coulter, Brea, CA, USA) were used for final library purification. Library quality was analyzed on a 2200 TapeStation System with Agilent High Sensitivity DNA 1000 ScreenTape System (Agilent Technologies, Santa Clara, CA, USA). Resulting libraries were subjected to paired-end Illumina sequencing (NovaSeq 6000, PE150; Illumina, San Diego, CA, USA).