title: GSM2061386: H9 ESC RNA-seq; Homo sapiens; RNA-Seq
library_construction_protocol: The mRNAs were purified using the Oligotex mRNA Min Kit (QIAGEN), according to the manufacturer’s instructions. After DNase treatment, 20 _g of total RNA were heated at 70◦C for 3 min to disrupt RNA secondary structure; the RNA was then incubated withOligotex suspension at 30◦C for 10 min. Oligotex-bound mRNAs were then centrifuged, washed and eluted with buffers provided in the kit. Total RNA (10 mg) was harvested from ESCs using TRI reagent (Ambion) for cDNA library preparation. Enrichment of mRNA by depletion of ribosomal RNA was performed using a RiboMinus transcriptome isolation kit (Invitrogen). RiboMinus RNA (1 mg) was then fragmented using RNase III for 10 min and cleaned up using a RiboMinus concentration module (Invitrogen).
identifiers.namespace: GEO
identifiers.id: GSM2061386