GSM9277065: [S. pombe] [Flag-Swi6 ChIP] [wt swi6 (1195)] [replicate C]; Schizosaccharomyces pombe; ChIP-Seq
600million cells were pelleted and resuspended in room temperature 1XPBS. Cells were fixed in 1.5 mM EGS (final) for 30 minutes at room temperature, followed by 1% formaldehyde (final) for 30 minutes at room temperature. Next glycine was added to a final concentration of 0.25M, incubated 5 minutes at room temperature. Pellets washed twice with cold 1XTBS. Lysate was spun down, pellet was isolated and flash frozen in liquid nitrogen. Libraries were prepared using NEBNext Mulitplex Oligos for Illumina (E7335L, Lot 10172541; #E6442S, Lot #10236110; and #E6440S Lot #10188082) and Ultra II FS DNA Library Prep Kit for Illumina (E7805L, Lot 10202083) following the manufacturer's protocol, “Protocol for FS DNA Library Prep Kit (E7805, E6177) with Inputs </=100 ng (NEB)”. 200 pg of input and IP DNA was used as starting material. Libraries were amplified using 11 cycles via PCR. NEBNext sample purification beads (E6178S, Lot 10185312) were used to clean up PCR-enriched adaptor-ligated DNA. DNA concentration was quantified using a Qubit4 Fluorometer and quality was checked by a 4200 TapeStation instrument (Agilent). Libraries were pooled to equimolar quantities for sequencing.