Chromatin Immunoprecipitation
Revision as of 15:23, 20 January 2016 by Davebridges (Talk | contribs) (→Buffers and Solutions Needed: made subsections)
Revision as of 15:23, 20 January 2016 by Davebridges (Talk | contribs) (→Buffers and Solutions Needed: made subsections)
This protocol is modified from the Myer's Lab ChIPseq protocol v011014 found here. The original citation for this methodology is:
Johnson DS, Mortazavi A, Myers RM, Wold B. Genome-Wide Mapping of in Vivo Protein-DNA Interactions. Science (80- ) 316: 1497–1502, 2007. doi:10.1126/science.1141319
Buffers and Solutions Needed
- 20% Formaldehyde (from 37% formaldehyde Sigma F87750)
- 2.5M Glycine
- PBS (cold)
- Farnham Lysis Buffer (cold)
- RIPA Buffer (cold)
- Dynabeads (Invitrogen cat#)
- PBS with 5 mg/mL BSA (cold)
- [LiCl Wash Buffer]] (cold)
- TE: 10 mM Tris 7.5, 0.1 mM EDTA (cold)
- ChIP Elution Buffer
Equipment
- Cool microfuge and swinging bucket centrifuge down to 4C