Rapid preparation of covalently closed circular DNA by acridine yellow affinity chromatography

Walter S. Vincent, Elliott S. Goldstein

Research output: Contribution to journalArticle

23 Scopus citations

Abstract

Covalently closed circular DNA can be isolated rapidly from cell lysates in a two-step process. Hydroxylapatite chromatography to prepurify the plasmid DNA from contaminating protein and RNA is followed by a step gradient elution of covalently closed circular (CCC) plasmid DNA from an acridine yellow affinity column. This procedure results in CCC DNA of a purity comparable to that obtained from ethidium bromide-CsCl gradients without lengthy centrifugation and free of contaimination by intercalating dye. Up to 250 μg of CCC pBR 322 can be isolated from 500 ml of bacterial culture in 4-6 h.

Original languageEnglish (US)
Pages (from-to)123-127
Number of pages5
JournalAnalytical Biochemistry
Volume110
Issue number1
DOIs
StatePublished - Jan 1 1981

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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