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 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'''. | # 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'''. | ||
''Turn water bath to 65°C before next step'' | |||
# 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. | ||