The Myc Gene pp 117-133

Part of the Methods in Molecular Biology book series (MIMB, volume 1012)

Chromatin Immunoprecipitation Assays for Myc and N-Myc

  • Bonnie L. Barrilleaux
  • Rebecca Cotterman
  • Paul S. Knoepfler
Protocol

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.

Key words

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

Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  • Bonnie L. Barrilleaux
    • 1
  • Rebecca Cotterman
    • 1
  • Paul S. Knoepfler
    • 1
  1. 1.Department of Cell Biology and Human Anatomy, Institute of Pediatric Regenerative MedicineUniversity of California Davis School of Medicine, Shriners Hospital For Children Northern CaliforniaSacramentoUSA

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