915
edits
Changes
Wrote initial section with buffers and link to new methods paper
[[ Category: ChIP ]]
[[ Category: RNA ]]
[[ Category: Transcription ]]
[[ Category: Molecular Biology ]]
This protocol is modified from the Myer's Lab ChIPseq protocol v011014 found [https://www.encodeproject.org/documents/6ecd8240-a351-479b-9de6-f09ca3702ac3/@@download/attachment/ChIP-seq_Protocol_v011014.pdf 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. [http://dx.doi.org/10.1126/science.1141319 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]]
[[ Category: RNA ]]
[[ Category: Transcription ]]
[[ Category: Molecular Biology ]]
This protocol is modified from the Myer's Lab ChIPseq protocol v011014 found [https://www.encodeproject.org/documents/6ecd8240-a351-479b-9de6-f09ca3702ac3/@@download/attachment/ChIP-seq_Protocol_v011014.pdf 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. [http://dx.doi.org/10.1126/science.1141319 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]]