GSM9531893: 105C cells, ENT, replicate 1; Homo sapiens; RNA-Seq
Total RNA was extracted with the RNeasy Kit (Qiagen, #74106) Total RNA was quantified and its integrity was assessed using 5K / RNA / Charge Variant Assay LabChip and RNA Assay Reagent Kit (Perkin Elmer). Libraries were generated from 250 ng of total RNA as follows: mRNA enrichment and library preparation were performed using the Illumina Stranded mRNA Prep Kit (Illumina), as per the manufacturers recommendations. Libraries were quantified using the KAPA Library Quantification Kits - Complete kit (Universal) (Kapa Biosystems). Average fragment size was determined using a Fragment Analyzer 5300 (Agilent) instrument. The libraries were normalized and pooled and then denatured in 0.02N NaOH and neutralized using pre-load buffer. The pool was loaded at 150pM on an Illumina NovaSeq X Plus 25B lane following the manufacturer's recommendations. The run was performed for 2x100 cycles (paired-end mode). A phiX library was used as a control and mixed with libraries at 1% level. Program BCL Convert 4.2.4 was then used to demultiplex samples and generate fastq reads.