RNA-SEQ of Silene linneana Voroschilov
Whole specimen plants were ground to a fine powder using mortar and pestle chilled on N2(l). RNA was extracted from 100 mg powder using the SpectrumTM Plant Total RNA Kit (Sigma Aldrich, St. Louis, MO) according to manufacturer's instructions utilising "protocol A", simultaneously employing on-column DNase I digestion. Mortars and pestles used for grinding frozen material were decontaminated between samples using 10% Sodium Hypochlorite (Prince 1992). One microgram of total RNA was used as input for sequencing library construction using TruSeqTM reagents (Illumina, San Diego, CA) according to manufacturers instructions. Samples were indexed with 12 multiplex adapters and sequenced with v3 clustering and SBS reagents, employing 100 bp paired end reads on 2 lanes of an HiSeq 2000 (Illumina) at the Norwegian Sequencing Center in Oslo, Norway (http://www.sequencing.uio.no).