RNAseq of Contarinia nasturtii: midgut rep 2
Total RNA was extracted separately from each tissue type and replicate using the standard TRIzol Reagent protocol (Invitrogen), and then further cleaned using an Illustra RNAspin Mini Kit with DNase treatment (GE Healthcare Life Sciences) to remove any salts and potential contaminants. Total RNA concentration was quantified using a NanoDrop One spectrophotometer (Thermo Scientific) and quality assessed on an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). Total RNA (1 g) from each replicate and tissue type was sent to the National Research Council of Canada (Saskatoon, SK) for RNA-Seq library preparation and sequencing. Libraries were prepared with the Illumina TruSeq stranded mRNA kit (Illumina, San Diego, CA) according to the manufacturers protocol. Paired-end, 125 bp cDNA libraries were sequenced on an Illumina HiSeq 2500 in a single lane, resulting in ~27 million reads per library.