title: GSM4699521: Gene expression data for SW480 monoculture, replicate 3; Homo sapiens; RNA-Seq
library_construction_protocol: At least 250 000 cells were collected by centrifugation and lysed using ExtractRNA (Evrogen, Russia). The cell lysates were stored at – 70°C until RNA extraction. RNA was extracted with RNA Clean & ConcentratorTM-5 (Zymo Research, United States). DNase I treatment was performed in-column manner during the clean-up. The quality of the extracted RNA was determined using TapeStation 2200 (Agilent Technologies, Santa Clara, California, United States) with Agilent High Sensitivity RNA ScreenTape (Agilent Technologies, United States). Since the RINe was greater than 7 for all samples, poly(A) RNA isolation was performed with a NEBNext Poly(A) mRNA Magnetic Isolation Module (New England BioLabs, Ipswich, Massachusetts, United States). The NEBNext® Ultra II Directional RNA Library Prep Kit for Illumina® with Sample Purification Beads (New England BioLabs, United States) and a NEBNext® Multiplex Oligos for Illumina (Index Primers Set 1-3) (New England BioLabs, United States) were used for the preparation of single-indexed libraries. After the preparation, the individually indexed libraries were mixed in equimolar amounts; the final mixture was analyzed with TapeStation using Agilent High Sensitivity DNA ScreenTape (Agilent Technologies, United States). The sequencing was performed with an Illumina NovaSeq 6000 System (Illumina, SP-type, single-end reads, 100 bp).