Chromatin immunoprecipitation assays for Myc and N-Myc

Bonnie L. Barrilleaux, Rebecca Cotterman, Paul S. Knoepfler

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Myc and N-Myc have widespread impacts on the chromatin state within cells, both in a gene-specific and genome-wide manner. Our laboratory uses functional genomic methods including chromatin immunoprecipitation (ChIP), ChIP-chip, and, more recently, ChIP-seq to analyze the binding and genomic location of Myc. In this chapter, we describe an effective ChIP protocol using specific validated antibodies to Myc and N-Myc. We discuss the application of this protocol to several types of stem and cancer cells, with a focus on aspects of sample preparation prior to library preparation that are critical for successful Myc ChIP assays. Key variables are discussed and include the starting quantity of cells or tissue, lysis and sonication conditions, the quantity and quality of antibody used, and the identification of reliable target genes for ChIP validation.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
Pages117-133
Number of pages17
Volume1012
DOIs
StatePublished - 2013

Publication series

NameMethods in Molecular Biology
Volume1012
ISSN (Print)10643745

Keywords

  • ChIP-seq
  • Chromatin immunoprecipitation
  • Epigenetics
  • Histone modifications
  • Myc
  • N-Myc

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Barrilleaux, B. L., Cotterman, R., & Knoepfler, P. S. (2013). Chromatin immunoprecipitation assays for Myc and N-Myc. In Methods in Molecular Biology (Vol. 1012, pp. 117-133). (Methods in Molecular Biology; Vol. 1012). https://doi.org/10.1007/978-1-62703-429-6-9