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

G Manfioletti; C Schneider

doi:10.1093/nar/16.7.2873

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

G Manfioletti·C Schneider

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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.

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Anion Exchange Resins Bacteriophage lambda Base Sequence Cetrimonium Cetrimonium Compounds Chromatography, Ion Exchange Cloning, Molecular DNA, Viral Detergents Ethanolamines Genetic Vectors Journal Article Research Support, Non-U.S. Gov't

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G Manfioletti, C Schneider. 1988-04-11. A new and fast method for preparing high quality lambda DNA suitable for sequencing.. https://doi.org/10.1093/nar%2F16.7.2873

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