GSM8900370: liver, TCPOBOP + SHAM, replicate 2; Mus musculus; RNA-Seq
Mouse livers were isolated 9h after surgery, thus 3h post-injection, and stored in RNA later. RNA was extracted using the Aurum Total RNA Mini kit (Bio-Rad) according to the manufacturer's-protocol Per sample, an amount of 1000ng of total RNA was used as input. Using the Illumina Stranded mRNA Sample Prep Kit (protocol version: # 1000000124518 v03, June 2022), poly-A containing mRNA molecules were purified from the total RNA input using poly-dT oligo-attached magnetic beads. The purified mRNA was fragmented, and in a reverse transcription reaction using random primers and Actinomycin D, RNA was converted into first strand cDNA, and subsequently into double-stranded cDNA in a second strand cDNA synthesis reaction using dUTP to achieve strand specificity. The cDNA fragments were extended with a single 'A' base to the 3' ends of the blunt-ended cDNA fragments, after which pre-index anchors were ligated preparing the fragments for dual indexing. Anchor-ligated fragments were then purified using magnetic beads. Finally, enrichment PCR was carried out to enrich those DNA fragments that have anchor-ligated DNA fragments and to add indexes and primer sequences for cluster generation. Purified dual-indexed sequence-libraries of each sample were equimolarly pooled and sequenced on Element Biosciences AVITI (2x75 Cloudbreak High kit, single read 100 (101-10-10-0)) at the VIB Nucleomics Core
GEO
GSM8900370_r1