Trueseq stranded rna seq library prep kit

Total RNA was isolated from five independent biological replicates of each infected and non-infected group using TRIzol reagent (Life Technologies, Carlsbad, CA, USA) according to the manufacturer’s instructions and as previously described^{39 (link)}. The RNA samples were treated with DNase I (1 U per µg of RNA) (Thermo Scientific, Lithuania, EU) at 37 °C for 1 h, and the RNA concentration was determined from the A260/A280 ratio using a NanoDrop ND1000 (Thermo Scientific, USA). In addition, RNA integrity was evaluated using an Agilent 2100 Bioanalyzer and a Pico Agilent RNA 6000 kit (Agilent Technologies, Santa Clara, CA, USA) according to the manufacturer’s instructions. rRNA depletion was performed using a poly(A) magnetic bead capture protocol and a TrueSeq Stranded Total RNA Sample Prep kit (Illumina) according to the manufacturer´s instructions. Libraries were prepared using a TrueSeq Stranded RNA-seq Library Prep Kit (Illumina), according to the manufacturer’s instructions.

Total RNA from five independent biological replicates was isolated using TRIzol reagent (Life Technologies) according to the manufacturer’s instructions. RNA samples were treated with DNase I (1 U per 1 µg of RNA) (Thermo Scientific) at 37 °C for 1 h, and the RNA concentration was determined using a spectrophotometer Nanodrop ND1000 spectrophotometer (Thermo Scientific). In addition, RNA integrity was evaluated using an Agilent 2100 Bioanalyzer and a Pico Agilent RNA 6000 kit (Agilent Technologies) according to the manufacturers’ instructions. rRNA depletion was performed using a poly (A) magnetic beads capture protocol and a TrueSeq Stranded Total RNA Sample Prep kit (Illumina) according to the manufacturers’ instructions. Libraries were prepared using a TrueSeq Stranded RNA-seq Library Prep kit (Illumina), according to the manufacturers’ instructions.