Influence of residual uracil-DNA glycosylase activity on the electrophoretic migration of dUTP-containing PCR products

Citation
M. Ritzler et al., Influence of residual uracil-DNA glycosylase activity on the electrophoretic migration of dUTP-containing PCR products, J MICROB M, 35(1), 1999, pp. 73-76
Citations number
9
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biology,Microbiology
Journal title
JOURNAL OF MICROBIOLOGICAL METHODS
ISSN journal
0167-7012 → ACNP
Volume
35
Issue
1
Year of publication
1999
Pages
73 - 76
Database
ISI
SICI code
0167-7012(199902)35:1<73:IORUGA>2.0.ZU;2-8
Abstract
In diagnostic amplification protocols contamination by previously amplified nucleic acids is considered the major source of false positive results. Su bstituting dUTP for dTTP in the PCR and initial treatment with uracil-DNA g lycosylase (UNG) virtually eliminates these carryover contaminations. Subse quent procedures to visualize the amplicons or to optimize sensitivity and specificity of the test are not always fully compatible with UNG-treated PC R products. Here we describe the more pronounced influence of residual UNG activity on the migration of PCR amplification products in polyacrylamide a s compared to agarose gels. (C) 1999 Elsevier Science B.V. All rights reser ved.