GSM8535024: RS4-11 cells, Control rep2; Homo sapiens; RNA-Seq
Cells were harvested and total RNA was isolated using the NucleoSpin RNA kit (Machery-Nagel). NGS library prep was performed with Illumina's Stranded mRNA Prep Ligation Kit following Stranded mRNA Prep Ligation ReferenceGuide (June 2020) (Document # 1000000124518 v00). Libraries were prepared with a starting amount of 1000 ng and amplified in 9 PCR cycles. After PCR purification, an additional round of purification was performed using 1x of AMPure XP beads. Libraries were profiled in a High Sensitivity DNA on a 2100 Bioanalyzer (Agilent technologies) and quantified using the Qubit dsDNA HS Assay Kit, in a Qubit 2.0 Fluorometer (Life technologies). All 9 samples (three replicates of untreated HEL cells and three replicates of MERCK60 treated and SIAH2 KO cells) were pooled in equimolar ratio and sequenced on 1 NextSeq500 high output FC, SR for 1x79 cycles plus 10 cycles for the index read and 2 dark cycle upfront R1.