GSM3589892: WT_137; Rattus norvegicus; RNA-Seq
RNA was extracted using the RNeasy Mini Kit (Qiagen), according to the manufacturer's instructions. Subsequently, the quantity of all purified RNA samples was measured on a nanodrop (2.07±0.01 A260/280; 2.11±0.19 A260/230) and the quality and integrity were measured with the Agilent 2100 Bioanalyzer (Agilent, Santa Clara, CA, USA). All RNA integrity numbers were greater than 9 (9.6±0.3). 1µg of total RNA was used for the preparation of the RNAseq library using the Illumina Genome Analyzer IIx TruSeq mRNA Seq Kit supplied by Illumina (Cat number: RS-122-2001). A poly-A-based mRNA enrichment step was carried out and cDNA was synthesized and used for library preparation using the Illumina TruSeqTM RNA sample preparation kit as previously described (Tariq MA et al. 2011), except for the following step: adapter-ligated DNA fragments were size-selected by gel-free size selection using appropriate concentration of SPRI AMPure beads to get an average 200bp peak size in adaptor ligated DNA. The size selected adaptor-ligated DNA fragments were amplified by LM-PCR. Then, Illumina recommended 6bp barcode bases were introduced at one end of the adaptors during PCR amplification step. The amplified PCR products were then purified with SPRI AMPure XP magnetic beads to get the final RNAseq library, which was used for high-throughput RNAseq. All samples were sequenced on the Illumina Genome Analyzer IIx.