title: GSM8881533: 145S; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA that was isolated from fresh frozen BC tumor and BM tissues using the mirVana™ PARIS™ RNA and Native Protein Purification Kit (Thermo Fisher Scientific, Waltham, MA, USA), following the manufacturer's instructions. Isolated RNA concentration and its purity were assessed by NanoDropTM 2000 spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA) determining the A260/280 and A260/230 ratios, respectively. RNA integrity number (RIN) and distribution value 200 (DV200) were assessed with an Agilent RNA 6000 Nano Kit on a 2100 Bioanalyzer (Agilent, Santa Clara, CA, USA). The RNA isolates were stored at -80° C. Next, 10 ng of RNA was used to prepare the RNA libraries. During the library preparation, rRNA was depleted using KAPA RNA HyperPrep with RiboErase and KAPA Globin Depletion Hybridization Oligos (Roche, Basel, Switzerland). SMART-Seq Stranded Kit (Takara Bio, Kusatsu, Japan) was used for low input RNA quantities. The libraries were sequenced on the NovaSeq 6000 platform (Illumina, San Diego, CA, USA). Depths of 30 million (6Gb) paired-end 100 bp reads were generated for each sample.