Phospho rpb1 ctd ser5 and ser2 antibody

CUT&Tag libraries (two biological replicates) were prepared using the CUT&Tag-IT^™ Assay Kit (Active Motif, 53160) following to the manufacturer’s protocol. 10⁶ of Mel cells were collected for each biological replicate and two replicates were prepared. The Mel cells were bound to Concanavalin A Beads and Incubated with 1:50 rabbit polyclonal Phospho-Rpb1 CTD Ser5 and Ser2 antibody (Cellsignaling 13523 and 13499). Guinea pig a-rabbit antibody was used at 1:100 dilution as secondary antibody. Tagmentation was performed using pA-Tn5 Transposomes at 37°C for 60 mins. DNA was purified by DNA Purification Column, then universal i5 primers and uniquely barcoded i7 primers were added to the DNA with 14cycles of PCR. Individual libraries were purified with SPRI beads and eluted with 20 µL DNA Purification Buffer. The libraries were sequenced on a MiSeq. For data analysis, see Supplementary Materials.

CUT&Tag libraries were prepared using the CUT&Tag-IT Assay Kit (Active Motif, 53160) following to the manufacturer’s protocol. 10⁶ of MEL cells were collected for each biological replicate and two replicates were prepared. The MEL cells were bound to Concanavalin A Beads and Incubated with 1:50 rabbit polyclonal Phospho-Rpb1 CTD Ser5 and Ser2 antibody (Cell Signaling 13523 and 13499). Guinea pig a-rabbit antibody was used at 1:100 dilution as secondary antibody. Tagmentation was performed using pA-Tn5 Transposomes at 37°C for 60 minutes. DNA was purified by DNA Purification Column, then universal i5 primers and uniquely barcoded i7 primers were added to the DNA with 14 cycles of PCR. Individual libraries were purified with SPRI beads and eluted with 20 μL DNA Purification Buffer. The libraries were sequenced on a MiSeq. Antibodies are listed in Table S1.