Separation of genomic DNA from plasmid DNA by selective renaturation with immobilized metal affinity capture

Tony Cano, Jason C. Murphy, George E. Fox, Richard C. Willson

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

In contrast to proteins, many nucleic acids can undergo reversible modification of their conformations, and this flexibility can be used to facilitate purification. Selective renaturation with capture is a novel method of removing contaminating genomic DNA from plasmid samples. Plasmid DNA quickly renatures after thermal denaturation and cooling (or alkaline denaturation followed by neutralization), whereas genomic DNA remains locally denatured after rapid cooling in mismatch-stabilizing high ionic strength buffer. Partially denatured genomic DNA can be selectively bound to a metal chelate affinity adsorbent through exposed purine bases, while double-stranded renatured plasmid DNA is not bound. Using this method we have readily achieved 1,000,000-fold clearance of 71 wt % contaminating E. coli genomic DNA from plasmid samples.

Original languageEnglish (US)
Pages (from-to)1472-1477
Number of pages6
JournalBiotechnology Progress
Volume21
Issue number5
DOIs
StatePublished - Sep 1 2005

ASJC Scopus subject areas

  • Biotechnology

Fingerprint Dive into the research topics of 'Separation of genomic DNA from plasmid DNA by selective renaturation with immobilized metal affinity capture'. Together they form a unique fingerprint.

Cite this