GSM7993758: GLV, model, rep2; Rattus norvegicus; RNA-Seq
Total RNA was extracted using Trizol (Invitrogen, Carlsbad, CA), and purified by chloroform/isoamyl alcohol extraction and 75% ethanol precipitation. 1 μg of total RNA was enriched for the Poly-A mRNA and reverse transcribed to double-stranded cDNA. RNA quality was detected by Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA). The qualified RNA samples are denatured at suitable temperature, and the mRNA is enriched by oligo (dT) -attached magnetic beads. The target mRNA fragments were reverse transcribed into cDNA, and the complementary cDNA chain was synthesized. The cDNA fragments were purified with the QiaQuick PCR extraction kit and eluted with EB buffer. The purified cDNA was end-repaired and the poly(A) were added. Suitable fragments were selected, purified, and subsequently PCR-amplified to establish cDNA library template. The prepared DNA library was sequenced on the Illumina HiSeq2500 platform.