title: GSM1419019: 3hH89 replicate2; Mus musculus; RNA-Seq
library_construction_protocol: RNA was isolated using Trizol (Invitrogen) according to manufacturer'instructures. 10 μg of total RNA was subjected to two rounds of polyA selection with Dynabeads® mRNA Purification kit (Invitrogen). mRNA was subsequently fragmented by hydrolysis (40 mM TrisOAc pH 8.2, 100 mM KOAc, 150 mM MgOAc) at 94oC for 3 min. 1st strand cDNA synthesis was performed using Superscript III Reverse Transcriptase kit (Invitrogen) with random hexamers priming (Applied Biosystems) in the presence of Actinomycin D (5 ng/ml).  Second strand cDNA was synthesized using DNA Pol I, DNA ligase (both Invitrogen) and RNase H (NEB) with random hexamers (Applied Biosystems) in the presence of dUTP. The libraries were prepared by CSF NGS unit (csf.ac.at) using NEBNext® Library Prep Reagent Set for Illumina (NEB).The following TrueSeq barcodes were used: sample 0h_1 TAGCTT, sample 0h_2 ATCACG, sample 1hA_1 GGCTAC, sample 1hA_2 CGATGT.
attributes.tag: GEO Accession
attributes.value: GSM1419019