A new and fast method for preparing high quality lambda DNA suitable for sequencing.
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.
Manfioletti
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G
European Molecular Biology Laboratory, Heidelberg, FRG.
Schneider
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C
D000837:Anion Exchange Resins; D010582:Bacteriophage lambda / Q000235:genetics*; D001483:Base Sequence; D000077286:Cetrimonium; D002593:Cetrimonium Compounds; D002852:Chromatography, Ion Exchange / Q000379:methods; D003001:Cloning, Molecular / Q000379:methods*; D004279:DNA, Viral / Q000302:isolation & purification*; D003902:Detergents; D004983:Ethanolamines; D005822:Genetic Vectors
D000837:Anion Exchange Resins; D002593:Cetrimonium Compounds; D004279:DNA, Viral; D003902:Detergents; D004983:Ethanolamines; C007369:2-diethylaminoethanol; D000077286:Cetrimonium