GSM4058820: SCT 31; Homo sapiens; RNA-Seq
Total RNA was extracted by double extraction protocol: RNA isolation by acid guanidinium thiocyanate-phenolchloroform extraction (TRIzol, Thermo Fisher Scientific, Waltham, MA, USA), followed by Qiagen RNeasy Micro clean-up (Qiagen, Hilden, Germany). cDNA libraries were prepared with 2 ng total RNA input and 1 μl of a 1:50,000 dilution of ERCC RNA Spike-in Controls (Ambion®, Thermo Fisher Scientific, Waltham, MA, USA) using the SMARTSeq v2 protocol with the following modifications: 1) 20 μM template switch oligos, and 2) 250 pg cDNA with 1/5 reaction of Illumina Nextera XT kit (Illumina, San Diego, CA, USA). The length distribution of the cDNA libraries was monitored using a DNA High Sensitivity Reagent Kit on Perkin Elmer Labchip GX system (Perkin Elmer, Waltham, MA, USA). All samples were subjected to an indexed PE sequencing run of 2x151 cycles on Illumina HiSeq 4000 platform (30 samples/lane).
GEO Accession
GSM4058820