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TitleEvaluation of uncertainty in quantitative real-time PCR
Publication TypeJournal Article
Year of Publication2006
AuthorsLove, J.L., Scholes P., Gilpin B., Savill M., Lin S., and Samuel L.
JournalJournal of Microbiological Methods
Volume67
Issue2
Pagination349 - 356
Date Published2006
ISSN01677012 (ISSN)
KeywordsAbsorption, analytic method, article, Calibration, Campylobacter jejuni, Chemistry Techniques, Analytical, Chemistry, Analytical, confidence interval, controlled study, Data analysis, DNA, DNA content, DNA determination, DNA, Bacterial, evaluation, measurement, nonhuman, nucleic acid, nucleotide sequence, Polymerase Chain Reaction, prediction, priority journal, quantitative analysis, real time polymerase chain reaction, reliability, Reproducibility of Results, sample, Taq Polymerase, uncertainty
AbstractQuantitative real-time PCR is one of the newer methods for measurement of the amount of nucleic material in biological systems. However, reliable measurement requires an appropriate estimation of uncertainty and this paper has developed the uncertainty budget associated with this procedure using as an example, data from a quantitative real-time PCR method for the enumeration of Campylobacter jejuni. This uncertainty is relatively large and for instance, a measured result of 151 units of DNA would have a 95% confidence interval of ± 84 units of DNA with the main sources of uncertainty being the measurement of the threshold cycle (Ct) value, the predicted DNA content of the unknown sample from the calibration line and the molar absorbance value for DNA. © 2006 Elsevier B.V. All rights reserved.
URLhttp://www.scopus.com/inward/record.url?eid=2-s2.0-33748744620&partnerID=40&md5=63563d03c96c33b9384abc73da234427
DOI10.1016/j.mimet.2006.04.005

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