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→Incubate bead-antibody complex with fragmented, cross-linked chromatin
==== Incubate bead-antibody complex with fragmented, cross-linked chromatin====
# Add 100 μl of antibody-coupled beads from above to each 25ug chromatin of preparation (from Sonication protocol) and incubate on a rotator for '''1 hour at room temperature''', followed by '''1 hour at 4°C'''.
# Collect beads containing immuno-bound chromatin by placing the microfuge tube on a magnet rack.
# Remove and discard supernatant. ''Turn water bath to 65°C before next step''.
# Wash beads 5 times with 1 mL LiCl Wash Buffer, mixing 3 minutes for each wash on a rotator.
# Add 1 ml TE Buffer to beads. Mix 1 minute on rotator and then place tubes on magnet rack to collect beads and discard supernatant.