Chromatin immunoprecipitation for studying transcriptional regulation in Xenopus oocytes and tadpoles.

David Stewart, Akihiro Tomita, Yun Bo Shi, Jiemin Wong

Research output: Contribution to journalReview article

13 Citations (Scopus)

Abstract

Understanding the accurate temporal and spatial regulation of gene expression during development requires knowledge of the spectrum of transcription factors and cofactors involved and their functional interplay with chromatin. Chromatin immunoprecipitation (ChIP) has become a powerful technique that allows us to do so. A typical ChIP assay involves (1) treating cells or tissues with formaldehyde to rapidly crosslink chromatin-associated proteins to DNA, (2) shearing chromatin by sonication into small fragments, (3) immunoprecipitation of the proteins of interest, (4) reversal of crosslinking, and (5) quantitating the specific associated DNA sequences by PCR. Here we present and discuss the protocols we have developed over the years for ChIP assays using Xenopus oocytes and tadpole tissues as experimental materials.

Original languageEnglish
Pages (from-to)165-181
Number of pages17
JournalMethods in molecular biology (Clifton, N.J.)
Volume322
Publication statusPublished - 01-01-2006
Externally publishedYes

Fingerprint

Chromatin Immunoprecipitation
Xenopus
Chromatin
Oocytes
Larva
Sonication
Gene Expression Regulation
Immunoprecipitation
Formaldehyde
Proteins
Transcription Factors
Polymerase Chain Reaction
DNA

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Cite this

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abstract = "Understanding the accurate temporal and spatial regulation of gene expression during development requires knowledge of the spectrum of transcription factors and cofactors involved and their functional interplay with chromatin. Chromatin immunoprecipitation (ChIP) has become a powerful technique that allows us to do so. A typical ChIP assay involves (1) treating cells or tissues with formaldehyde to rapidly crosslink chromatin-associated proteins to DNA, (2) shearing chromatin by sonication into small fragments, (3) immunoprecipitation of the proteins of interest, (4) reversal of crosslinking, and (5) quantitating the specific associated DNA sequences by PCR. Here we present and discuss the protocols we have developed over the years for ChIP assays using Xenopus oocytes and tadpole tissues as experimental materials.",
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Chromatin immunoprecipitation for studying transcriptional regulation in Xenopus oocytes and tadpoles. / Stewart, David; Tomita, Akihiro; Shi, Yun Bo; Wong, Jiemin.

In: Methods in molecular biology (Clifton, N.J.), Vol. 322, 01.01.2006, p. 165-181.

Research output: Contribution to journalReview article

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