ArticleBMC biotechnology2005
Standardisation of data from real-time quantitative PCR methods - evaluation of outliers and comparison of calibration curves.
Article in BMC biotechnology, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 76 papers.
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Who cites it
76 citing papers in PubMed.
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- Quantitative Real-Time PCR for Trypanosoma cruzi Infection Diagnosis and Treatment Response Monitoring of Patients with Chagas Disease.Methods in molecular biology (Clifton, N.J.) · 2026Article
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- PEGylated Multimeric RNA Nanoparticles for siRNA Delivery in Traumatic Brain Injury.Small (Weinheim an der Bergstrasse, Germany) · 2025Article
- Differentially expressed lncRNAs in SOD1Open biology · 2024Article
- A Standard Framework for Evaluating Large Health Care Data and Related Resources.MMWR supplements · 2024Article
- C-C motif chemokine ligand 2 regulates prostaglandin synthesis and embryo attachment of the bovine endometrium during implantation.Cell and tissue research · 2024Article
- Myeloid Cell Association with Spinal Cord Injury-Induced Neuropathic Pain and Depressive-like Behaviors in LysM-eGFP Mice.The journal of pain · 2024Article
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- Generation of isogenic models of Angelman syndrome and Prader-Willi syndrome in CRISPR/Cas9-engineered human embryonic stem cells.PloS one · 2024Article
- Transcription of microRNAs is regulated by developmental signaling pathways and transcription factors.Frontiers in cell and developmental biology · 2024Article
- Activation of inflammasomes and their effects on neuroinflammation at the microelectrode-tissue interface in intracortical implants.Biomaterials · 2023Article
16 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundAs real-time quantitative PCR (RT-QPCR) is increasingly being relied upon for the enforcement of legislation and regulations dependent upon the trace detection of DNA, focus has increased on the quality issues related to the technique. Recent work has focused on the identification of factors that contribute towards significant measurement uncertainty in the real-time quantitative PCR technique, through investigation of the experimental design and operating procedure. However, measurement uncertainty contributions made during the data analysis procedure have not been studied in detail. This paper presents two additional approaches for standardising data analysis through the novel application of statistical methods to RT-QPCR, in order to minimise potential uncertainty in results.
resultsExperimental data was generated in order to develop the two aspects of data handling and analysis that can contribute towards measurement uncertainty in results. This paper describes preliminary aspects in standardising data through the application of statistical techniques to the area of RT-QPCR. The first aspect concerns the statistical identification and subsequent handling of outlying values arising from RT-QPCR, and discusses the implementation of ISO guidelines in relation to acceptance or rejection of outlying values. The second aspect relates to the development of an objective statistical test for the comparison of calibration curves.
conclusionThe preliminary statistical tests for outlying values and comparisons between calibration curves can be applied using basic functions found in standard spreadsheet software. These two aspects emphasise that the comparability of results arising from RT-QPCR needs further refinement and development at the data-handling phase. The implementation of standardised approaches to data analysis should further help minimise variation due to subjective judgements. The aspects described in this paper will help contribute towards the development of a set of best practice guidelines regarding standardising handling and interpretation of data arising from RT-QPCR experiments.
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