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Basic principles of real-time quantitative PCR
Abstract
Real-time quantitative PCR allows the sensitive, specific and reproducible quantitation of nucleic acids. Since its introduction, real-time quantitative PCR has revolutionized the field of molecular diagnostics and the technique is being used in a rapidly expanding number of applications. This exciting technology has enabled the shift of molecular diagnostics toward a high-throughput, automated technology with lower turnaround times. This article reviews the basic principles of real-time PCR and describes the various chemistries available: the double-stranded DNA-intercalating agent SYBR Green 1, hydrolysis probes, dual hybridization probes, molecular beacons and scorpion probes. Quantitation methods are discussed in addition to the competing instruments available on the market. Examples of applications of this important and versatile technique are provided throughout the review.
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Citations
Cited by
Quan, P.-L., Sauzade, M., & Brouzes, E. (2018) · Sensors · doi:10.3390/s18041271 · Cited by 664
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Afzal, A. (2020) · Journal of Advanced Research · doi:10.1016/j.jare.2020.08.002 · Cited by 363
Viruses in wastewater: occurrence, abundance and detection methods
Corpuz, M. V. A., Buonerba, A., Vigliotta, G., Zarra, T., Ballesteros, F., Campiglia, P., Belgiorno, V., Korshin, G., & Naddeo, V. (2020) · Science of The Total Environment · doi:10.1016/j.scitotenv.2020.140910 · Cited by 325
Scaling up: A guide to high‐throughput genomic approaches for biodiversity analysis
Porter, T. M., & Hajibabaei, M. (2018) · Molecular Ecology · doi:10.1111/mec.14478 · Cited by 305
CRISPR/cas systems redefine nucleic acid detection: Principles and methods
Wang, M., Zhang, R., & Li, J. (2020) · Biosensors and Bioelectronics · doi:10.1016/j.bios.2020.112430 · Cited by 268
Metadata
- Authors
- Arya, M., Shergill, I. S., Williamson, M., Gommersall, L., Arya, N., & Patel, H. R.
- Venue
- Expert Review of Molecular Diagnostics
- Publisher
- Informa UK Limited
- Published
- 2005-03
- Volume
- 5
- Issue
- 2
- Pages
- 209-219
- ISSN
- 1473-7159 (ISSN-L)
- 1744-8352
- OpenAlex
- W2141439851
- Cited by
- 627
- References
- 0
- FWCI
- 7.03
- Type
- journal article
- Language
- en
- Open access
- closed
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- Arya, M., Shergill, I. S., Williamson, M., Gommersall, L., Arya, N., & Patel, H. R. (2005). Basic principles of real-time quantitative PCR. Expert Review of Molecular Diagnostics. https://doi.org/10.1586/14737159.5.2.209
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