Changes

Jump to: navigation, search

Chromatin Immunoprecipitation

73 bytes added, 18:00, 26 January 2016
Crosslinking, Lysis and Shearing of DNA
2. Add formaldehyde to a final concentration of 1% directly to the media of adherent cells growing on tissue culture plates, swirl gently, and
incubate at room temperature for 10 minutes.
* If using 10cm dishes add 270ul of 37% formaldehyde
3. Stop the cross-linking reaction by adding glycine to a final concentration of 0.125M (and swirl gently to mix.* If using 10cm dishes add 0.5mL of the 2.5M glycine stock solution) and swirl gently to mix. 
4. Remove media from plates and wash cells with equal volume cold (4°C) 1X PBS.
* 10mL for 10cm dish
5. Aspirate the PBS and add 2-3 .5 ml cold (4°C) Farnham lysis buffer.
6. Scrape the cells off the plate with a cell scraper and transfer into 15-ml conical tubes on ice.
162
edits

Navigation menu