Illumina HiSeq 4000 paired end sequencing; RNA-seq of microbial communities from Namib Desert gravel plain soils, dry and watered
RNA-seq of microbial communities from Namib Desert gravel plain soils, dry and watered
Approximately 2x 10 g of surface soil (excluding pebbles) was embedded in RNAlater solution (Sigma) and frozen. 30 L/m2 water with \"Namib desert salts\" solution added at time zero. Dry control samples were left untreated. Soil was washed in 5 volumes of ice-cold 10 mM Tris-HCl 1 mM EDTA pH 6.5 buffer containing 100 mM NaH2PO4 to remove RNAlater salts. CTAB lysis buffer (5% CTAB, 0.7 M NaCl, 240 mM KH2PO4, pH 8). TRIzol (Sigma) RNA extraction following recommended protocols. Purified RNA was treated with DNAseI and re-precipitated with LiCl. ScriptSeq Complete Gold Kit (epidemiology) (Illumina)