GSM5013442: CD8 T cells normal rabbit serrum CUT&RUN; Mus musculus; OTHER
CD8 T cells were isolated from mouse spleeen using anti-mouse CD8a magnetic beads (BD 55151).  50M were isolated for Chip and kept frozen (from 5 mice) and 150,000 were isolated from one mouse and freshly used for CUT&RUN.  For ChIP: cells were thawed, lyzed and sonicated followed by ChIP with rabbit anti -Runx3 serum (1:200) and capturing of DNA-Ab complexes using protein-A sepharose magnetic beads. For CUT&RUN, cells were immobilised on ConA magnetic beads, permeabilized and incubated with rabbit anti-Runx3 serum (1:200) and procedure followed according to (Skene P J  2017 eLife e21856). Following deproteinization, DNA fragments were used for library construction through the protocol NEBNEXT Ultra II DNA library prep (E7645) with 12 cycles of PCR amplification. Samples were sequenced on Illumina Next-seq 500 machine.