title: GSM4763715: Macrophages_ACP+TNF_24 hours [TNF.PRP_rep3]; Homo sapiens; RNA-Seq
library_construction_protocol: RNA was extracted using the RNeasy mini kit (Qiagen) from co-cultures of macrophages and fibroblasts (control, ACP, TNF, or ACP+TNF for 24h; n=4/ea). A total of 100 ng of high-quality RNA (RIN>8 and A260/280>1.8) were used to construct libraries. RNA-Seq library preparations were done using established Illumina methods for mRNA-Seq (Part #RS-122-2001). Briefly, poly A+ RNA was purified from 100 ng of total RNA with oligo-dT beads and fragmented with divalent cations at elevated temperature. First strand cDNA synthesis was performed with random hexamer priming and SuperScript II Reverse transcriptase (Invitrogen, part # 18064-014). Second strand cDNA synthesis was performed using RNAseH and DNA PolI. Following dscDNA synthesis, the double stranded products are end repaired, followed by addition of a single 'A' base and then ligation of the Illumina TruSeq adaptors. The resulting product was amplified with 15 cycles of PCR. Each library was made with a unique Index sequence and libraries are pooled for sequencing.
attributes.tag: GEO Accession
attributes.value: GSM4763715