GSM5017798: PHF10_total_exp1_2; Homo sapiens; RNA-Seq
RNA was isolated from 3 millions of cell with TriReagent (MRC) according to the manufacturer's protocol. Total RNA from A375 cells after knockdown of MYC or PHF10 (isolated as described above) was treated with DNase I (Thermo Fisher Sci.) at 370C for 15 min followed by phenol-chloroform extraction and 96% ethanol precipitation. For NGS library preparation, one µg of RNA and NEBNext Ultra II Directional RNA library prep kit for Illumina with NEBNext Poly(A) mRNA magnetic isolation module (NEB) were used according to the manufacturer's protocol. Adaptor ligation was performed using NEBNext multiplex oligos for Illumina index primer set #1 (NEB). Quality of RNA and cDNA libraries was analyzed with Bioanalyzer RNA 6000 Nano Chip and Bioanalyzer DNA 1000 Chip kits (Agilent). RNA and DNA yields were assessed with Qubit assay kits (Thermo Fisher Sci.). Libraries were sequenced on an Illumina NovaSeq 6000 with single strand 100 bp length reads. Approximately 3x107 reads were obtained per one sample.