Analysis of distant communication on defined chromatin templates in vitro

Methods Mol Biol. 2009:543:563-76. doi: 10.1007/978-1-60327-015-1_33.

Abstract

Regulation of many biological processes in eukaryotes involves distant communication between the regulatory DNA sequences (e.g., enhancers) and their targets over the DNA regions organized in chromatin. However previously developed methods for analysis of communication in chromatin in vitro are artifact-prone and/or do not allow analysis of communication on physiologically relevant, saturated arrays of nucleosomes. Here we describe a method for quantitative analysis of the rate of distant communication in cis on saturated arrays of nucleosomes capable of forming the 30-nm chromatin fibers in vitro.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Buffers
  • Chickens
  • Chromatin / metabolism*
  • Chromatin Assembly and Disassembly
  • DNA / isolation & purification
  • DNA Primers / metabolism
  • DNA Restriction Enzymes / metabolism
  • Electrophoresis
  • Enhancer Elements, Genetic / genetics
  • Indicators and Reagents
  • Molecular Biology / methods*
  • Nucleosomes / metabolism
  • Promoter Regions, Genetic / genetics
  • RNA / isolation & purification
  • Templates, Genetic
  • Transcription, Genetic

Substances

  • Buffers
  • Chromatin
  • DNA Primers
  • Indicators and Reagents
  • Nucleosomes
  • RNA
  • DNA
  • DNA Restriction Enzymes