期刊
BIOTECHNOLOGY LETTERS
卷 24, 期 24, 页码 2053-2056出版社
KLUWER ACADEMIC PUBL
DOI: 10.1023/A:1021319421153
关键词
gene quantification; PCR efficiency; primers; RT-PCR
The effect of primer selection on real-time polymerase chain reaction (RT-PCR) performance was tested. Primer sets of varying length of product were used to amplify the sequence of beta-actin. Variability in length caused variability in RT-PCR performance. Kinetic parameters of PCR were studied by mathematical approximation of real-time data by means of a four-parametric sigmoid model. This model describes the full kinetics of the amplification trajectory. Statistical exploration of parameters yielded by this model revealed that reactions with higher amplification efficiency - primed by well-performing primers - proceed with lower variability and are therefore better suited for measurement purposes.
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