title: GSM1355147: Otx2-/-_tetON_Otx2_plusdox; Mus musculus; RNA-Seq
library_construction_protocol: RNAs from two independent biological replicates of ESC culture and two independent biological replicates from EpiLCs were extracted with Trizol (Invitrogen), following the manufacturer's recommendations. 10 μg of total RNA were subjected to two rounds of oligo-dT purification using Dynaloligo-dT beads (Invitrogen). 100 ng of the purified RNA were fragmented with 10x fragmentation buffer (Ambion). Fragmented RNA was used for first-strand cDNA synthesis, using random hexamer primers (Invitrogen) and SuperScript II enzyme (Invitrogen). Second strand cDNA was obtained by adding RNaseH (Invitrogen) and DNA Pol I (New England Biolabs) to the first strand cDNA mix. The resulting double-stranded cDNA was used for Illumina library preparation and sequenced with Illumina Genome Analyzer.
attributes.tag: GEO Accession
attributes.value: GSM1355147