Effects of primer-template mismatches on the polymerase chain reaction: human immunodeficiency virus type 1 model studies.
Nucleic Acids Res
; 18(4): 999-1005, 1990 Feb 25.
Article
en En
| MEDLINE
| ID: mdl-2179874
ABSTRACT
We investigated the effects of various primer-template mismatches on DNA amplification of an HIV-1 gag region by the polymerase chain reaction (PCR). Single internal mismatches had no significant effect on PCR product yield while those at the 3'-terminal base had varied effects. AG, GA, and CC mismatches reduced overall PCR product yield about 100-fold, AA mismatches about 20-fold. All other 3'-terminal mismatches were efficiently amplified, although the GG mismatches appeared to be more sensitive to sequence context and dNTP concentrations than other mismatches. It should be noted that mismatches of T with either G, C, or T had a minimal effect on PCR product yield. Double mismatches within the last four bases of a primer-template duplex where one of the mismatches is at the 3' terminal nucleotide, in general, reduced PCR product yield dramatically. The presence of a mismatched T at the 3'-terminus, however, allowed significant amplification even when coupled with an adjacent mismatch. Furthermore, even two mismatched Ts at the 3'-terminus allowed efficient amplification.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Composición de Base
/
Productos del Gen gag
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Reacción en Cadena de la Polimerasa
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VIH-1
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Técnicas de Amplificación de Ácido Nucleico
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Genes Virales
Idioma:
En
Revista:
Nucleic Acids Res
Año:
1990
Tipo del documento:
Article
País de afiliación:
Canadá