title: GSM3639298: P6_2h; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was harvested from the BMSCs using the TRIzol reagent (Invitrogen) according to the manufacturer's protocol. The total RNA was treated with the RQ1 DNase (Promega) to remove DNA. The quality and quantity of the purified RNA were determined by measuring the absorbance at 260 nm/280 nm (A260/A280) using the SmartsSpec Plus spectrophotometer (BioRad). The RNA integrity was further verified by 1.5% agarose gel electrophoresis. For each sample, 5μg of total RNA was used for RNA-seq library preparation. Polyadenylated mRNAs were purified and concentrated with oligo(dT)-conjugated magnetic beads (Invitrogen). The purified mRNAs were fragmented at 95 ℃, followed by end repair and 5' adaptor ligation. Then RNA reverse transcription was performed with an RT primer harbouring a 3' adaptor sequence and randomized hexamers. The cDNAs were purified and amplified with the RNA-Seq Library Preparation Kit (Gnomegen). PCR products corresponding to 200-500 bps were purified, quantified and stored at -80 ℃ before sequencing.
attributes.tag: GEO Accession
attributes.value: GSM3639298