Protocol

Xenopus Protocols

Volume 322 of the series Methods in Molecular Biology™ pp 139-147

Chromatin Assembly of DNA Templates Microinjected Into Xenopus Oocytes

  • Danièle RocheAffiliated withResearch Section, Institut Curie, UMR218 du Centre National de la Recherche Scientifique (CNRS)
  • , Geneviève AlmouzniAffiliated withResearch Section, Institut Curie, UMR218 du Centre National de la Recherche Scientifique (CNRS)
  • , Jean-Pierre QuivyAffiliated withResearch Section, Institut Curie, UMR218 du Centre National de la Recherche Scientifique (CNRS)

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Abstract

The packaging of deoxyribonucleic acid (DNA) into chromatin within the eukaryotic nucleus can affect processes such as DNA replication, transcription, recombination, and repair. Therefore, studies aimed at understanding at the molecular level how these processes are operating have to take into account the chromatin context. We present a method to assemble DNA into chromatin by nuclear microinjection into Xenopus oocytes. This method allows in vivo chromatin formation in a nuclear environment. We provide the experimental procedures for oocyte preparation, DNA injection, and analysis of the assembled chromatin.

Key Words

Microinjection oocytes chromatin assembly nucleosomes