GSM8285637: iPS_BasalCnl_TTseq_rep1; Homo sapiens; OTHER
After treatment, cells were harvested and lysed with Trizol. ERCC spike-ins (00043, 00170, 00136) were added to lysates after which RNA was isolated, fragmented and biotinylated with EZ-link HPDP-biotin (Thermo Fischer. Biotinylated nascent RNAs were purified with streptavidin-conjugated magnetic beads (µMACS; Miltenyi) and libraries of total and biotinylated RNA were prepared with Illumina TruSeq total RNA kit. Libraries were quantified using the KAPA Library Quantification Kit and sequenced with Illumina NextSeq 500 using the High Output Kit v2.5 (150 cycles, Illumina) for 2 × 75 bp paired-end reads. Libraries of biotinylated RNA were prepared with Illumina TruSeq total RNA kit. Libraries were quantified using the KAPA Library Quantification Kit and sequenced with Illumina NextSeq 500 using the High Output Kit v2.5 (150 cycles, Illumina) for 2 × 75 bp paired-end reads.