title: GSM3439695: FD41488 P35 SRC-2/NCOA2 siRNA day 3 EPC; Homo sapiens; RNA-Seq
library_construction_protocol: RNeasy Plus kit (Qiagen Inc., Germantown, MD) 250ng of total RNA as input, the TruSeqTM RNA library preparation kit v2 (Illumina®, Inc., San Diego, CA) was used for mRNA isolation, fragmentation, and subsequent first and second strand DNA synthesis.  The kit also was used for follow up end-repair along with base- and adaptor- ligation before products were purified and then enriched by PCR (15 cycles) to create the cDNA libraries.  The resulting libraries were quantitated using the NanoDropTM spectrometer and fragment size assessed using a DNA 1000 chip on the Agilent 2100 Bioanalyzer. A qPCR assay was performed on the libraries to determine the concentration of adapter-ligated fragments using the Applied Biosystems ViiATM 7 Real-Time PCR System (Thermo Fisher Scientific®) with a KAPA library Quant kit (Kapabiosystems, Inc., Wilmington, MA). Relevant samples were pooled in equimolar amounts and re-quantitated by qPCR. Library pools (27pM) were loaded into a high output flow cell for clonal cluster generation by bridge amplification using the cBot 2 system (Illumina®, Inc.). Indexed paired-end sequencing (2x100bp read lengths) was performed using the ultrahigh-throughput HiSeq 2500 sequencing system (Illumina®, Inc.).
attributes.tag: GEO Accession
attributes.value: GSM3439695