alias: GSM4715544
title: GSM4715544: SLC22A15_OE_A1; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted from the above clones using E.Z.N.A Total RNA kit I (OMEGA bio-tek) following the manufacturer's protocol. Strand-specific RNA-seq libraries were prepared using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina following the manufacturer's recommendations. In summary, total RNA quality was assessed using an RNA 6000 Nano kit on an Agilent 2100 Bioanalyzer (Agilent Biotechnologies), and the concentration was measured using a Qubit RNA HS assay kit (Life Technologies). A 500 ng aliquot of total RNA was depleted of rRNA using a Human/Mouse/Rat RNAse-H based Depletion kit from NEB (New England Biolabs, Ipswich, MA). Following rRNA removal, mRNA was fragmented and then used for first- and second-strand cDNA synthesis with random hexamer primers. Double-stranded cDNA fragments underwent end repair, poly (A) tailing and the ligation of two unique adapters (Integrated DNA Technologies, Coralville, IA). The adaptor-ligated cDNA was amplified by limited-cycle PCR. Library quality was analyzed with TapeStation High-Sensitivity D1000 ScreenTape (Agilent Biotechnologies) and quantified by KAPA qPCR (KAPA BioSystems). Libraries were pooled and sequenced at 2 × 150-bp read lengths on the Illumina HiSeq 4000 platform.
identifiers.namespace: GEO
identifiers.id: GSM4715544
attributes.tag: GEO Accession
attributes.value: GSM4715544