Journal
BMC BIOINFORMATICS
Volume 8, Issue -, Pages -Publisher
BMC
DOI: 10.1186/1471-2105-8-85
Keywords
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Background: Reverse transcription followed by real-time PCR is widely used for quantification of specific mRNA, and with the use of double-stranded DNA binding dyes it is becoming a standard for microarray data validation. Despite the kinetic information generated by real-time PCR, most popular analysis methods assume constant amplification efficiency among samples, introducing strong biases when amplification efficiencies are not the same. Results: We present here a new mathematical model based on the classic exponential description of the PCR, but modeling amplification efficiency as a sigmoidal function of the product yield. The model was validated with experimental results and used for the development of a new method for real-time PCR data analysis. This model based method for real- time PCR data analysis showed the best accuracy and precision compared with previous methods when used for quantification of in-silico generated and experimental real- time PCR results. Moreover, the method is suitable for the analyses of samples with similar or dissimilar amplification efficiency. Conclusion: The presented method showed the best accuracy and precision. Moreover, it does not depend on calibration curves, making it ideal for fully automated high-throughput applications.
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