A new and fast method for preparing high quality lambda DNA suitable for sequencing

Nucleic Acids Res. 1988 Apr 11;16(7):2873-84. doi: 10.1093/nar/16.7.2873.

Abstract

A method is described for the rapid purification of high quality lambda DNA. The method can be used from either liquid or plate lysates and on a small scale or a large scale. It relies on the preadsobtion of all polyanions present in the lysate to an "insoluble" anion-exchange matrix (DEAE or TEAE). Phage particles are then disrupted by combined treatment with EDTA/proteinase K and the resulting DNA is precipitated by the addition of the cationic detergent cetyl (or hexadecyl)-trimethyl ammonium bromide-CTAB ("soluble" anion-exchange matrix). The precipitated CTAB-DNA complex is then exchanged to Na-DNA and ethanol precipitated. The resultant purified DNA is suitable for enzymatic reactions and provides a high quality template for dideoxy-sequence analysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anion Exchange Resins
  • Bacteriophage lambda / genetics*
  • Base Sequence
  • Cetrimonium
  • Cetrimonium Compounds
  • Chromatography, Ion Exchange / methods
  • Cloning, Molecular / methods*
  • DNA, Viral / isolation & purification*
  • Detergents
  • Ethanolamines
  • Genetic Vectors*

Substances

  • Anion Exchange Resins
  • Cetrimonium Compounds
  • DNA, Viral
  • Detergents
  • Ethanolamines
  • 2-diethylaminoethanol
  • Cetrimonium