GSM5140823: RML1000 (Lung 5 D21); Mus musculus; RNA-Seq
Total RNA from lung tissue samples from different groups was extracted following the double extraction protocol: RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction followed by a Qiagen RNeasy Micro clean-up procedure. Total RNAs were analyzed on Agilent Bioanalyser for quality assessment with RNA Integrity Number (RIN) range from 4.2 to 6.7 and median of RIN 5.1. Ribosomal RNA (rRNA) were depleted from 500ng of RNA using Ribo-Zero™ Gold (Human/Mouse/Rat) according to manufacturer's protocol. cDNA libraries were prepared from the resultant rRNA depleted RNA using Lexogen SENSE Total RNA-Seq Library Prep Kit according to manufacturer's protocol for degraded RNA except with 21 PCR cycles and an additional cleanup with 0.9X Ampure XP beads. The length distribution of the cDNA libraries was monitored using a DNA High Sensitivity Reagent Kit on the Perkin Elmer Labchip. All samples were subjected to an indexed paired-end sequencing run of 2x151 cycles on an Illumina HiSeq 4000 system.