Evaluation of PCR-based methods for isolating flanking regions of genes.
Mol Biotechnol
; 32(2): 111-6, 2006 Feb.
Article
en En
| MEDLINE
| ID: mdl-16444012
ABSTRACT
Several polymerase chain reaction (PCR)-based methods are available for isolation of unknown genomic fragments. In the present study, a comparative evaluation of a few methods of ligation-mediated PCR methods and a ligation-independent one were made by isolating promoter fragment for N-methyltransferase gene involved in the caffeine biosynthetic pathway of Coffea canephora. The benefits of tertiary PCR and the effects of a 4-base cutting restriction endonuclease on the size of the PCR products obtained were demonstrated in one of the ligation-mediated PCR methods. The methods adopted in this study differed in the sizes of the 5'-flanking regions obtained. The efficiencies of various methods used reflect the inherent limitations of the PCR-based methods for isolation of unknown flanking regions.
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Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Biotecnología
/
Reacción en Cadena de la Polimerasa
/
ADN de Plantas
/
Región de Flanqueo 5'
Tipo de estudio:
Diagnostic_studies
/
Evaluation_studies
Idioma:
En
Revista:
Mol Biotechnol
Asunto de la revista:
BIOLOGIA MOLECULAR
/
BIOTECNOLOGIA
Año:
2006
Tipo del documento:
Article
País de afiliación:
India