Construction of plasmids containing site-specific DNA interstrand cross-links for biochemical and cell biological studies

Methods Mol Biol. 2012:920:203-19. doi: 10.1007/978-1-61779-998-3_15.

Abstract

Plasmids containing a site-specific DNA interstrand cross-link (ICL) are invaluable tools for the investigation of ICL repair pathways at the biochemical and cellular level. We describe a procedure for preparation of plasmid DNA substrates containing a single ICL at a specific site. The procedure is versatile, leads to reliable yields of pure DNA substrate, and is suitable for the incorporation of virtually any type of DNA lesion into plasmids.

Publication types

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

MeSH terms

  • Base Sequence
  • Cisplatin / pharmacology
  • Cross-Linking Reagents / pharmacology*
  • DNA / chemistry*
  • DNA / genetics
  • DNA / metabolism*
  • DNA Adducts / metabolism
  • DNA Repair / drug effects
  • DNA Restriction Enzymes / metabolism
  • Plasmids / chemistry*
  • Plasmids / metabolism*

Substances

  • Cross-Linking Reagents
  • DNA Adducts
  • DNA
  • DNA Restriction Enzymes
  • Cisplatin