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Open Access Software

FCI: an R-based algorithm for evaluating uncertainty of absolute real-time PCR quantification

Paolo Verderio1, Sara Pizzamiglio1, Fabio Gallo1 and Simon C Ramsden2*

Author Affiliations

1 Unit of Medical Statistics and Biometry, Istituto Nazionale dei Tumori, Milan, Italy

2 National Genetics Reference Laboratory (Manchester), St. Mary's Hospital, Manchester, UK

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BMC Bioinformatics 2008, 9:13  doi:10.1186/1471-2105-9-13

Published: 10 January 2008

Abstract

Background

FCI is an R code for analyzing data from real-time PCR experiments. This algorithm estimates standard curve features as well as nucleic acid concentrations and confidence intervals according to Fieller's theorem.

Results

In order to describe the features of FCI four situations were selected from real data collected during an international external quality assessment program for quantitative assays based on real-time PCR. The code generates a diagnostic figure suitable for assessing the quality of the quantification process.

Conclusion

We have provided a freeware programme using this algorithm specifically designed to increase the information content of the real-time PCR assay.