Chromatin Immunoprecipitation Cloning Protocol

- from the Farnham lab. Click here for full protocol link.

Day 1

1.  Add formaldehyde directly to tissue culture media to a final concentration of 1%.  We generally use 1 x 107 cells per antibody per timepoint.  For cloning, multiple IPs are performed and pooled at later steps therefore we generally start with 1 x 108 cells.  Incubate adherent cells on a shaking platform and suspension cells on a stir plate for 10 minutes at room temperature. 

2.  Stop the crosslinking reaction by adding glycine to a final concentration of 0.125 M.  Continue to rock or spin at room temp for 5 minutes.