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

Authors :
Claudio Schneider
Guidalberto Manfioletti
Manfioletti, Guidalberto
Schneider, C.
Source :
Nucleic Acids Research. 16:2873-2884
Publication Year :
1988
Publisher :
Oxford University Press (OUP), 1988.

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.

Details

ISSN :
13624962 and 03051048
Volume :
16
Database :
OpenAIRE
Journal :
Nucleic Acids Research
Accession number :
edsair.doi.dedup.....7a010d935bd3439ba93c2c8ebbada439
Full Text :
https://doi.org/10.1093/nar/16.7.2873