ArticleNucleic acids research2003
Mathematics of quantitative kinetic PCR and the application of standard curves.
Article in Nucleic acids research, 2003. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 183 papers.
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Who cites it
183 citing papers in PubMed, 530 citations in OpenAlex.
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- Quantification Revisited: What qPCR Efficiency Models Reveal About Data Analysis Integrity.International journal of molecular sciences · 2026Review
- Seed shattering in a North American Oryzeae grain: Developmental and genomic signatures of early domestication.The plant genome · 2026Article
- Development and application of a TaqMan probe-based pentaplex qPCR for the rapid detection and differential identification of five pathogens.Frontiers in microbiology · 2026Article
- Evaluation of culture- and PCR-based methods for detecting Burkholderia pseudomallei in soil samples in Thailand.PLoS neglected tropical diseases · 2026Article
- A novel TaqMan probe-based pentaplex qPCR assay for the simultaneous detection of five pathogenic protozoans.Scientific reports · 2025Article
- Species-level enumeration of low-concentration lactic acid bacteria in breast milk.Food chemistry. Molecular sciences · 2025Article
- Peptidoglycan-reshuffling proteins SCO0954, SCO1758, SCO4439, and SCO4440 modulate the formation of wall-deficient cells in Streptomyces coelicolor under hyperosmotic sucrose stress.Scientific reports · 2025Article
- Differences in the suitability of published honey bee (Apis mellifera) reference genes between the African subspecies Apis mellifera scutellata and European derived Apis mellifera.Molecular ecology · 2025Article
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- The DeoR-like pleiotropic regulator SCO1897 controls specialised metabolism, sporulation, spore germination, and phosphorus accumulation in Streptomyces coelicolor.Communications biology · 2024Article
- Development of a novel sequence based real-time PCR assay for specific and sensitive detection of Burkholderia pseudomallei in clinical and environmental matrices.Annals of clinical microbiology and antimicrobials · 2024Article
- Article
- Sperm function, mitochondrial activity and in vivo fertility are associated to their mitochondrial DNA content in pigs.Journal of animal science and biotechnology · 2024Article
- Quantitative real-time PCR assays for species-specific detection and quantification of Baltic Sea spring bloom dinoflagellates.Frontiers in microbiology · 2024Article
- Normalizing real-time PCR results in routine testing.Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc · 2024Article
- Development of a Novel Internally Controlled HrpB1 Gene-Based Real-Time qPCR Assay for Detection of Burkholderia pseudomallei.Molecular diagnosis & therapy · 2024Article
123 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Fluorescent monitoring of DNA amplification is the basis of real-time PCR, from which target DNA concentration can be determined from the fractional cycle at which a threshold amount of amplicon DNA is produced. Absolute quantification can be achieved using a standard curve constructed by amplifying known amounts of target DNA. In this study, the mathematics of quantitative PCR are examined in detail, from which several fundamental aspects of the threshold method and the application of standard curves are illustrated. The construction of five replicate standard curves for two pairs of nested primers was used to examine the reproducibility and degree of quantitative variation using SYBER Green I fluorescence. Based upon this analysis the application of a single, well- constructed standard curve could provide an estimated precision of +/-6-21%, depending on the number of cycles required to reach threshold. A simplified method for absolute quantification is also proposed, in which quantitative scale is determined by DNA mass at threshold.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.