title: GSM4497322: Brain, Cont13_1; Rattus norvegicus; RNA-Seq
library_construction_protocol: To construct an RNA-Seq library, we first isolated total cellular RNA from brain tissue samples using TRIzol reagent (Ambion, Austin, TX, USA) according to the manufacturer's protocol and then treated them with RQ1 DNase (Promega, Madison, WI, USA) to remove contaminated DNA and quantified them by measuring the absorbance at 260 /280 nm with the Smartspec plus (BioRad, Hercules, CA, USA). The integrity of the RNA samples was further assessed using 1.5% agarose gel electrophoresis. Ten μg of each total RNA sample was used to construct the RNA-seq library. Specifically, polyadenylated mRNA samples were isolated, purified, and concentrated with oligo (dT)-conjugated magnetic beads (Invitrogen, Carlsbad, CA, USA) and fragmented at 95oC and ligated by the end repair and 5' adaptor. Next, these mRNA samples were reversely transcribed into cDNA using the RT primer harboring 3'-end adaptor sequences and randomized hexamer, which was purified and amplified using PCR, leading to 200-500 bp PCR products, which was purified, quantified and stored at -80oC before DNA sequencing.
attributes.tag: GEO Accession
attributes.value: GSM4497322