A standard additions method reduces inhibitor-induced bias in quantitative real-time PCR.
Anal Bioanal Chem
; 401(10): 3221-7, 2011 Dec.
Article
en En
| MEDLINE
| ID: mdl-22002559
ABSTRACT
A method of calibration for real-time quantitative polymerase chain reaction (qPCR) experiments based on the method of standard additions combined with non-linear curve fitting is described. The method is tested by comparing the results of a traditionally calibrated qPCR experiment with the standard additions experiment in the presence of 2 mM EDTA, a known inhibitor chosen to provide an unambiguous test of the principle by inducing an approximately twofold bias in apparent copy number calculated using traditional calibration. The standard additions method is shown to substantially reduce inhibitor-induced bias in quantitative real-time qPCR.
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Base de datos:
MEDLINE
Asunto principal:
Reacción en Cadena en Tiempo Real de la Polimerasa
Idioma:
En
Revista:
Anal Bioanal Chem
Año:
2011
Tipo del documento:
Article
País de afiliación:
Reino Unido