title: GSM2897330: THP-1 macrophages stimulated with nBSA for 24 hours; Homo sapiens; RNA-Seq
library_construction_protocol: The total RNA were isolated with RNeasy mini kit (Qiagen, Germany) according to the manufacturer's instructions The libraries were prepared with KAPA Stranded mRNA-Seq Kit for Illumina (Kapa biosystems, USA) following the manufacturer's procedure. In Brief, high quality intact total RNA (200 ng in 50 µl RNase-free water) were input for Poly(A) RNA capture with magnetic oligo-dT beads. The RNA was fragmented to the desired size (about 300 bp) by heating in the presence of Mg++. The 1st strand cDNA was synthesized with random primers. The 2nd strand cDNA synthesis converted cDNA:RNA hybrid to double strand cDNA (dscDNA), while marking the 2nd strand with dUTP. Then dAMP was added to 3'-end of dscDNA fragments. Following by ligation of 3'-dTMP adapters (10 nM) to 3'-dAMP library fragments, and PCR amplification (14 cycles) of the adapter-ligated library DNA. The dUTP-marked strand was not amplified. Library fragment size distribution was confirmed by electrophoresis and library concentration was determined with Qubit 2.0 Fluorometric Quantitation (Thermo Fisher Scientific, USA). These libraries were sequenced using the Illumina Hiseq 3000 Sequencer (50 cycles, single read lane).
attributes.tag: GEO Accession
attributes.value: GSM2897330