GSM6602132: LPS and recombinant AAT [6104]; Mus musculus; RNA-Seq
Total RNA was isolated from five mice samples per experimental group and qualitatively assessed by Agilent 2100 Bioanalyzer using Agilent RNA 6000 Nano Kit. RNASeq libraries were prepared by using 200 ng RNA of each sample. TruSeq Stranded mRNA (Illumina Inc., USA) was used for library preparation according to the manufacturer protocol. Briefly, polyA-containing RNA molecules were purified using polyT oligo-attached magnetic beads followed by thermal fragmentation. The cleaved RNA fragments were copied into first strand cDNA using reverse transcriptase and random primers. This was followed by a second strand cDNA synthesis, end repair process, adenylation of 30 ends and ligation of the adapters. The products were then purified with Ampure XP Beads (Beckman Coulter, Brea, CA), and enriched with 15 cycles of PCR to create the cDNA library.