title: GSM2827737: ChA+ rep1; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted from each fractionated sample using TRIzol LS, and ribosomal RNA was depleted using the RiboMinus™ Transcriptome Isolation Kit according to manufacturer's instructions. Polyadenylated RNA was then selected by Oligo (dT)25 Dynabeads following the instructions provided by manufacturer. The flow through was collected, and the RNA was extracted using TRIzol LS as non-polyadenylated RNA. Three biological replicates (independent cell culture) were obtained for each fraction. To prepare for RNA-Seq libraries, 50ng of RNA was extracted from each sample. Paired-end strand-specific RNA-Seq was performed with the NEBNext Ultra Directional RNA Library Prep Kit with the following adjustments to the manufacturer's instructions: fragmentation was performed for 12 minutes at 94°C and PCR cycling was set to 12 cycles. Unique index was ligated to each sample RNA library construct using the NEBNext® Multiplex Oligos for Illumina® Index Primers Sets 1&2.
identifiers.namespace: GEO
identifiers.id: GSM2827737