Chromatin Immunoprecipitation: Difference between revisions
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Wrote initial section with buffers and link to new methods paper |
→Buffers and Solutions Needed: made subsections |
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Johnson DS, Mortazavi A, Myers RM, Wold B. Genome-Wide Mapping of in Vivo Protein-DNA Interactions. Science (80- ) 316: 1497–1502, 2007. [http://dx.doi.org/10.1126/science.1141319 doi:10.1126/science.1141319] | Johnson DS, Mortazavi A, Myers RM, Wold B. Genome-Wide Mapping of in Vivo Protein-DNA Interactions. Science (80- ) 316: 1497–1502, 2007. [http://dx.doi.org/10.1126/science.1141319 doi:10.1126/science.1141319] | ||
==Buffers and Solutions Needed== | ===Buffers and Solutions Needed=== | ||
* 20% Formaldehyde (from 37% formaldehyde Sigma F87750) | * 20% Formaldehyde (from 37% formaldehyde Sigma F87750) | ||
* 2.5M Glycine | * 2.5M Glycine | ||
| Line 19: | Line 19: | ||
* TE: 10 mM Tris 7.5, 0.1 mM EDTA (cold) | * TE: 10 mM Tris 7.5, 0.1 mM EDTA (cold) | ||
* [[ChIP Elution Buffer]] | * [[ChIP Elution Buffer]] | ||
===Equipment=== | |||
* Cool microfuge and swinging bucket centrifuge down to 4C | |||
Revision as of 15:23, 20 January 2016
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