Chromatin Immunoprecipitation: Difference between revisions

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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 step 1.i above) to each 300 μl chromatin preparation (from Sonication protocol) and incubate on a rotator for one hour at room temperature, followed by one hour at  4°C.
# Add 100 μl of antibody-coupled beads (from step 1.i above) to each 25ug chromatin of preparation (from Sonication protocol) and incubate on a rotator for '''one hour at room temperature''', followed by '''one hour at  4°C'''.
# Collect beads containing immuno-bound chromatin by placing the microfuge tube on a magnet rack.
# Collect beads containing immuno-bound chromatin by placing the microfuge tube on a magnet rack.
# Remove and discard supernatant.
# Remove and discard supernatant.