RNA-seq with metabolic labelling of human cell lines K562 and HEK293FT treated with vorinostat and untreated controls
RNA-seq with metabolic labelling of human cell lines K562 and HEK293FT treated with vorinostat and untreated controls
K562 cells: RPMI 1640 medium with 10% fetal bovine serum  and 1% Penicillin/Streptomycin at 37°C and 5% CO2, seeded to a density of 2*10^5 cells/ml and split 1:4 once the density approached 10^6 cells/ ml.  HEK293FT cells: complete DMEM medium with 4.5 g/l glucose, 6 mM L-glutamine, 10% fetal bovine serum, 0.1 mM MEM nonessential amino acids, 1 mM sodium pyruvate and 100 µg/ml Penicillin-Streptomycin, dissociated with TrypLE Express before re-seeding at 1:3 density every three to four days. During treatment, cells were kept in the incubator before they were spun down for 5 minutes at 300 RCF at the end of the incubation. RNA extraction was performed using the MGIEasy Nucleic Acid Extraction Kit. The sequencing libraries were prepared from isolated RNA through mRNA capture with an oligo-dT primer, and full length cDNA molecules were constructed through the use of a template switching oligo. The resulting cDNA fragments were pre-amplified through PCR and then fragmented through tagmentation to yield sequence ready fragments.