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A new and fast method for prearing high quality lambda DNA suitable for sequencing
- 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.
- Subjects :
- Lysis
DNA purification
Detergents
Genetic Vectors
Matrix (chemical analysis)
Bacteriophage
chemistry.chemical_compound
Bone plate
Genetics
DNA sequencing
Cloning, Molecular
Anion Exchange Resins
Cetrimonium bromide
Chromatography
Base Sequence
biology
Cetrimonium
Chromatography, Ion Exchange
Proteinase K
biology.organism_classification
Bacteriophage lambda
DNA extraction
chemistry
Biochemistry
Ethanolamines
DNA, Viral
Cetrimonium Compounds
biology.protein
DNA
Subjects
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