Open AccessA new and fast method for prearing high quality lambda DNA suitable for sequencing
Author(s) -
Guidalberto Manfioletti,
Claudio Schneider
Publication year - 1988
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/16.7.2873
Subject(s) - dna , lysis , bromide , ammonium bromide , biology , cationic polymerization , ion exchange , lambda phage , enzyme , proteinase k , chromatography , biochemistry , combinatorial chemistry , ion , materials science , pulmonary surfactant , inorganic chemistry , chemistry , escherichia coli , bacteriophage , organic chemistry , polymer chemistry , gene
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.