Illumina HiSeq 4000 sequencing; Chip-seq of S. cerevisiae to investigate post-transcriptional buffering of low nascent RNA transcription by Puf5
Chip-seq of S. cerevisiae to investigate post-transcriptional buffering of low nascent RNA transcription by Puf5
Samples were grown in 60 mL YPD to an OD595 of 0.6-0.8. To 50 mL culture, formaldehyde was added (1% final concentration) and incubated for 60 min at 20 °C. The samples were centrifuged and washed 2x with ice-cold PBS and the pellet snap frozen in liquid nitrogen. Chromatin immunoprecipitation was performed as described (Tessarz P, Santos-Rosa H, Robson SC, Sylvestersen KB, Nelson CJ, Nielsen ML, et al. Glutamine methylation in histone H2A is an RNA-polymerase-I-dedicated modification. Nature. 2014;505: 564–568) Library production was performed as described (Tessarz P, Santos-Rosa H, Robson SC, Sylvestersen KB, Nelson CJ, Nielsen ML, et al. Glutamine methylation in histone H2A is an RNA-polymerase-I-dedicated modification. Nature. 2014;505: 564–568)