Chromatin Immunoprecipitation: Difference between revisions
| Line 93: | Line 93: | ||
==== Reverse cross-linking and recover ChIP DNA ==== | ==== Reverse cross-linking and recover ChIP DNA ==== | ||
# Incubate bead pellet from step 2.f above in a 65°C water bath for 1 hour, shake or vortex every 15 minutes to elute the immuno-bound chromatin from the beads. | # Incubate bead pellet from step 2.f above in a 65°C water bath for '''1 hour''', shake or vortex every 15 minutes to elute the immuno-bound chromatin from the beads. | ||
# Spin at 14,000 rpm in a microfuge at room temperature for 3 minutes. | # Spin at 14,000 rpm in a microfuge at room temperature for 3 minutes. | ||
# Collect the supernatant, which contains the ChIP’d DNA. The tubes can be placed on the magnet to facilitate supernatant recovery. | # Collect the supernatant, which contains the ChIP’d DNA. The tubes can be placed on the magnet to facilitate supernatant recovery. | ||