title: GSM4491154: BC04; Homo sapiens; RNA-Seq
library_construction_protocol: RNA extraction and purification from FFPE tissue sections were carried out using the RNeasy FFPE Kit (Qiagen, cat. no. 73504). The RNeasy MinElute spin columns were incubated for 5 minutes with RNase-free water before centrifugation. Samples were eluted in 20uL RNase-free water. A DNase treatment was added using HL-dsDNase kit (ArticZymes, cat. no. 70800-201 250U), with additive MgCl2 (ThermoFisher Scientific, cat. no. AB0359) and DTT, Molecular Grade (Promega, cat. no. P117A). RNA samples were DNase-treated at room temperature for 15 minutes, followed by a 55°C 10-minute incubation to complete enzyme activity.  RNA was stored at -80°C. RNA samples were prepared with Ovation® Human FFPE RNA-Seq Library Systems (Nugen, cat. no. 0340, 0341). Clean-up steps, including magnetic bead purifications, were performed with freshly made 70% EtOH from 100% proof Absolute Ethanol (Fisher BioReagents™, cat. no. BP2818-500), nuclease-free water (Ambion®, cat. no. AM9939), and DynaMag™-96 Side magnetic plates (Invitrogen, cat. no. 12331D). As a rule, each library was amplified for 18 PCR cycles. Libraries were pooled in 8 sample sets, and sequenced in a paired-end mode for 75 cycles, Mid-Output platform on an Illumina NextSeq 2000 Sequencing System.
attributes.tag: GEO Accession
attributes.value: GSM4491154