GSM7717573: Caco cells without MEVs 1; Homo sapiens; RNA-Seq
RNA was extracted with Trizol methods followed by DNase treatment and RNA purification by RNeasy MinElute Cleanup Kit (Qiagen) total RNA was quantified and its integrity was assessed using a LabChip GXII (PerkinElmer) instrument. Libraries were generated from 250 ng of total RNA using Illumina® Stranded mRNA Prep, Ligation kit (Illumina), as per the manufacturer's recommendations. Libraries were quantified using the KAPA Library Quanitification Kits - Complete kit (Universal) (Kapa Biosystems). Average size fragment was determined using a LabChip GXII (PerkinElmer) instrument. The libraries were normalized and pooled and then denatured in 0.05N NaOH and neutralized using HT1 buffer. The pool was loaded at 175pM on an Illumina NovaSeq S4 lane using Xp protocol as per 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. Base calling was performed with RTA v3.4.4. Program bcl2fastq2 v2.20 was then used to demultiplex samples and generate fastq reads.