title: GSM2405995: IMR-32 cells with Negative Control siRNA; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was isolated using Trizol reagent (Ambion, Foster City, CA) and quantified with a ND-1000 spectrophotometer (Thermo Scientific, Wilmington, DE). The quality of Total RNA was assessed by the Agilent Bioanalyzer Nano chip (Agilent Technologies). RNA-Seq library was constructed using the Truseq Stranded Total RNA library preparation instruction from Illumina using 1ug of Total RNA. Shortly, the total RNA is Ribo-depleted using Ribo-Zero Gold kit to remove rRNA from total RNA and then fragmented into small pieces using divalent cations under elevated temperature. Following fragmentation, the first strand cDNA were synthesized using random primers and followed by second strand synthesis using DNA Polymerase I. The cDNA is then ligated with index adapters for each sample followed by purification and then enriched with PCR to create the final library. The quality and quantity of the libraries were detected by Agilent Bioanalyzer and Kapa Biosystems qPCR. Multiplexed libraries are pooled and single-end 50-bp sequencing was performed on one flow-cell of an Illumina Hiseq 1500.
identifiers.namespace: GEO
identifiers.id: GSM2405995