GSM8174193: KOLF2 cells, SETBP1_PATH3, Day 0, biol rep 2; Homo sapiens; RNA-Seq
Cells were collected on day 0 and 24 for RNA extraction using RNeasy (Thermo Fisher). gDNA for amplicon sequencing was extracted using PureLinkTM Genomic DNA Mini Kit (Life Technologies) RNA integrity was determined using the Agilent TapeStation system (Genomics WA). RNA sequencing was performed according to SureSelect Strand-Specific RNA Library preparation for Illumina Multiplexed sequencing, paired-end, 100 bp, 30 M read RNA sequencing on the NOVAseq 6000 platform (Illumina, USA) at Genomics WA (Perth, Australia). PCR amplicon products corresponding to the SNV sites in SETBP1, PATH2, PATH3 and VUS2, were prepared from extracted gDNA with forward and reverse amplicon primers and Phusion High-Fidelity PCR Master Mix with HF Buffer (Thermo Scientific). Purified samples were barcoded with TruSeq primers (IDT) and SETBP1 sequences determined with paired-end, 250bp DNA amplicon sequencing with a MiniSeq Mid-Output Kit, 300 cycles.