GSM5643355: Oxy_rep2; Homo sapiens; RNA-Seq
Cells were lysed in 700 μL of QIAzol Lysis Reagent (Qiagen, Hilden, Germany) with subsequent RNA extraction with miRNeasy Mini Kit (Qiagen) according to the manufacturer protocol, including on-column treatment with RNase-Free DNase Set (Qiagen). RNA was extracted in 30 μL of RNase-free water. RNA concentration was measured with NanoDrop 2000 Spectrophotometer (Thermo Fisher Scientific). A260/A280 and A260/A230 ratios were more than 2.0, and RNA Integrity Number (RIN) according to 2100 Bioanalyzer with Agilent RNA 6000 Pico Kit (Agilent Technologies, Santa Clara, CA, USA) was not less than 9.7 for all RNA samples. Libraries for mRNA sequencing were prepared with TruSeq Stranded mRNA Library Prep (Illumina, San Diego, CA, United States). Library quality check was provided with Agilent High Sensitivity DNA Kit (Agilent Technologies). Sequencing with NextSeq 550 (Illumina) generated single-end 75-nucleotide reads.