GSM6069619: I-miR-200c-3p_sorafenib_1; Homo sapiens; RNA-Seq
Total RNA was extracted using a miRNeasy mini kit with DNase treatment according to the manufacturer's instructions(QIAGEN, Germany). Libraries were prepared using total RNA lysates. Total RNA concentration and quality of the RNA were assessed with Qubit (Qubit™ DNA HS assay) and TapeStation 4150 with the RNA ScreenTape Assay  (Agilent Technologies Genomics, USA), respectively. RNA Integrity Number (RIN) values were > 9 in all the RNA samples. Libraries were constructed from 350 ng of RNA with the QuantSeq 3' mRNA-Seq Library Prep Kit FWD for Illumina with unique dual indexes (set B1), folowing manufacturer instructions (Catalog number 114, Lexogen GmbH, Viena, Austria). Libraries were amplified (cycles 15-19), purified and subjected to quality control with High Sensitivity DNA Qubit® Assay and High Sensitivity D100 ScreenTape Assay (average size 318 bp). Samples were then pooled together at 2 nM, diluted at 750 nM (PhiX 2%) and sequenced in P2  flow cells  in a NextSeq 2000 instrument from Illumina (65 cycles)