RNAseq of wildtype and deletion mutants (H3.V KO, H4.V KO, H3.V/H4.V KO)

RNA was extraced from wildtype and deletion mutants and a comparative analysis of gene expression was performed using DESeq2. Total RNA from 45 million cells was extracted in triplicates, spiked with a synthetic RNA control. RNA was depleted for rRNA using custom-made anti-rRNA oligonucleotides that hybridize to the target RNAs and are subsequently digested with RNAseH. RNA was reverse transcribed into cDNA and libraries for Illumina sequencing were constructed. We aimed at 15 million reads for each replicate. Comparative expression analysis was performed using DeSeq2.