ArticlePloS one2013
Accurate and precise DNA quantification in the presence of different amplification efficiencies using an improved Cy0 method.
Article in PloS one, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed.
- A new diagnostic kit using real-time polymerase chain reaction for the detection of Mycobacterium tuberculosis.AMB Express · 2026Article
- Improved protocol forExperimental biology and medicine (Maywood, N.J.) · 2023Article
- Data treatment methods for real-time colorimetric loop-mediated isothermal amplification reactions.Scientific reports · 2023Article
- Clinical Validation of a Colorimetric Loop-Mediated Isothermal Amplification Using a Portable Device for the Rapid Detection of SARS-CoV-2.Diagnostics (Basel, Switzerland) · 2023Article
- shinyCurves, a shiny web application to analyse multisource qPCR amplification data: a COVID-19 case study.BMC bioinformatics · 2021Article
- Genomic analysis of high copy-number sequences for the targeted detection of Listeria species using a flow-through surveillance system.Archives of microbiology · 2021Article
- PANDAA intentionally violates conventional qPCR design to enable durable, mismatch-agnostic detection of highly polymorphic pathogens.Communications biology · 2021Article
- Framework for DNA Quantification and Outlier Detection Using Multidimensional Standard Curves.Analytical chemistry · 2019Article
- A novel data processing method CyC* for quantitative real time polymerase chain reaction minimizes cumulative error.PloS one · 2019Article
- Transcriptome signature for dietary fructose-specific changes in rat renal cortex: A quantitative approach to physiological relevance.PloS one · 2018Article
- Article
- A survey of tools for the analysis of quantitative PCR (qPCR) data.Biomolecular detection and quantification · 2014Review
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Authors and funding
6 authors.
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Abstract
Quantitative real-time PCR represents a highly sensitive and powerful technology for the quantification of DNA. Although real-time PCR is well accepted as the gold standard in nucleic acid quantification, there is a largely unexplored area of experimental conditions that limit the application of the Ct method. As an alternative, our research team has recently proposed the Cy0 method, which can compensate for small amplification variations among the samples being compared. However, when there is a marked decrease in amplification efficiency, the Cy0 is impaired, hence determining reaction efficiency is essential to achieve a reliable quantification. The proposed improvement in Cy0 is based on the use of the kinetic parameters calculated in the curve inflection point to compensate for efficiency variations. Three experimental models were used: inhibition of primer extension, non-optimal primer annealing and a very small biological sample. In all these models, the improved Cy0 method increased quantification accuracy up to about 500% without affecting precision. Furthermore, the stability of this procedure was enhanced integrating it with the SOD method. In short, the improved Cy0 method represents a simple yet powerful approach for reliable DNA quantification even in the presence of marked efficiency variations.
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