Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook

Table of Contents

Expand All | Contract All

ChIPQuantitative PCR (ChIP-qPCR)

(Protocol summary only for purposes of this preview site)

It is critical to determine if the ChIP actually enriched the DNA sequences that are associated with the target protein. If there are known genomic binding sites, primers can be designed for quantitative PCR (qPCR) to determine if the known sites are specifically enriched by immunoprecipitation compared with the input chromatin and with the chromatin that has been mock-immunoprecipitated with preimmune serum. If there are no known sites but candidate target genes are available, one can consider designing qPCR primers along the length of potential regulatory regions such as promoters and conserved noncoding sequences within intergenic and genic regions. Because real-time PCR can be performed in either a 96- or 384-well format in a minimal reaction volume and primers can be synthesized with minimal cost, ChIP-qPCR is an attractive strategy to interrogate target genes and potential regulatory regions across a large number of experimental conditions and different cell types. If candidate target genes or potential sites are not available, ChIP-chip (Protocol 5) or ChIP-seq (Protocol 6) should be considered.


Previous   |   Next

Save 30% & Get Free Shipping!

Save 30% at checkout on our website.
(Limited time special offer.) Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook
Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook

Search for information about other protocols included in the book:

Read What Others Are Saying About Molecular Cloning:

Tom Maniatis wins Lasker

'Not Quite a Book Prize'

Get alerts about special offers from Cold Spring Harbor Laboratory Press free to your Inbox