GSM5016367: donor2_purified_RNA_calmidazolium; Homo sapiens; RNA-Seq
Total RNA was extracted and purified from cell cultures using RNeasy plus mini kit from QIAGEN (Hilden, Germany). For in-lysate RNA, cell cultures were washed with PBS, treated with a lysis buffer consisting of 0.1% (1mg/mL) BSA (New England Biolabs, Ipswich, MA, Item No. B9000S) / 0.3% Igepal CA 630 (Sigma-Aldrich, St. Louis, MO, Item No. I8896) for 5 minutes, and the supernatant was used for further library prep. Extracted RNA and cell lysate were used for preparing sequencing libraries according to the Smart-3SEQ protocol (Foley et al., 2019). Illumina TruSeq libraries were prepared at the University of Iowa Genomics Facility using standard Illumina TruSeq stranded mRNA protocols. Concentration and fragment size of prepared library was measured using Qubit HS and bioanalyzer. Fragment size for Smart-3SEQ library was ~500bp which falls within the recommended 200-600bp