title: GSM2551475: SNA_Saline rep3; Mus musculus; RNA-Seq
library_construction_protocol: RNA was extracted using RNeasy kit (Qiagen), according to manufacturer’s guidelines. Residual DNA contamination was removed by treating the spin column with 40 units of RNase-free DNase I (Qiagen) for 15 min at room temperature prior to RNA elution. RNA concentrations and purity were verified for each sample following elution with Agilent 2100 Bioanalyzer (Agilent). RNA with RIN factor above 7.5 was used for library preparation. Libraries were generated using the TruSeq stranded mRNA Sample Preparation Kit A (Illumina, San Diego CA) according to the recommended low-sample TruSeq RNA Sample Preparation Guide protocol. cDNA wasgenerated through reverse transcriptase PCR using Superscript II reverse transcriptase (Invitrogen). cDNA was bound for purification during the protocol with Agencourt AMPure XP beads (Beckman-Coulter). Following ligation of unique adapter sequences, the DNA was enriched by PCR with 15 cycles of amplification according to the TruSeq protocol. Ligation and library integrity was verified using a High Sensitivity DNA chip on the Agilent 2100 Bioanalyzer (Agilent) with clean elution profiles at the correct size peak of about 260 bp. The Illumina TruSeq kit “A” adapter sequences were ligated to each sample in each group to allow for sequencing multiplexing. Sample libraries ligated with unique adapter sequences were multiplexed six to a lane and were sequenced using Illumina HiSeq 2000 100-cycle, single end sequencing.
attributes.tag: GEO Accession
attributes.value: GSM2551475