title: GSM3731568: Teratoma_Control_rep3; Mus musculus; RNA-Seq
library_construction_protocol: Teratoma samples were homogenized with an Ultra-TURRAX disperser (IKA) in presence of 2 ml TRIzol reagent (Invitrogen) using approximately 200 mg tissue per tumor. After clearance of supernatant by centrifugation for 5 min at 12.000 x g at 4°C, 0.4 ml of chloroform was added and the solution centrifuged for 15 min at 12.000 x g at 4°C. RNA from aqueous phase was precipitated with 1 ml isopropanol, washed with 75 % ethanol, air dried and resuspended in RNase-free water. Contaminating DNA was digested with DNase I followed by an RNA cleanup (RNeasy kit, Qiagen) according to manufacturer's instructions. RNA was quantified with a Thermo NanoDrop and quality tested on Agilent Bioanalyzer. NGS libraries were prepared using the TruSeq RNA Sample Prep Kit v2 (Illumina) according to the manufacturer's recommendations and amplified in 12 PCR cycles. Libraries were profiled in a High Sensitivity DNA chip on a 2100 Bioanalyzer and quantified using the Qubit dsDNA HS Assay Kit, in a Qubit 2.0 Fluorometer (Life Technologies). The NGS libraries were sequenced on a HiSeq 2000 Illumina sequencer, for 51 cycles plus seven cycles for the index read.
attributes.tag: GEO Accession
attributes.value: GSM3731568