Post-hybridization recovery of membrane filter-bound DNA for enzymatic DNA amplification.
Biotechniques
; 14(4): 575-8, 1993 Apr.
Article
em En
| MEDLINE
| ID: mdl-8476600
ABSTRACT
We describe here a simple and rapid method for enzymatic DNA amplification using DNA template recovered from membrane filters previously used in hybridization analysis. This is done by first solubilizing membrane pieces carrying DNA of interest in dimethyl sulfoxide, followed by isopropanol precipitation and polymerase chain reaction amplification. The source of membrane-bound DNA successfully tested includes plasmid and human leukocyte DNA and DNA immobilized on bacterial colony filters and plaque lifts. The sensitivity of the procedure is such that DNA recovered from 0.5 microgram of filter-bound total human DNA was enough for PCR amplification of a 0.3-kb fragment. Our protocol will be useful for recycling of scarce DNA samples for cloning and sequencing purposes.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
DNA Bacteriano
/
Reação em Cadeia da Polimerase
Tipo de estudo:
Diagnostic_studies
/
Evaluation_studies
Idioma:
En
Revista:
Biotechniques
Ano de publicação:
1993
Tipo de documento:
Article
País de afiliação:
China