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Multiplex PCR: Critical Parameters and Step-by-Step Protocol
Open accessAbstract
By simultaneously amplifying more than one locus in the same reaction, multiplex PCR is becoming a rapid and convenient screening assay in both the clinical and the research laboratory. While numerous papers and manuals discuss in detail conditions influencing the quality of PCR in general, relatively little has been published about the important experimental factors and the common difficulties frequently encountered with multiplex PCR. We have examined various conditions of the multiplex PCR, using a large number of primer pairs. Especially important for a successful multiplex PCR assay are the relative concentrations of the primers at the various loci, the concentration of the PCR buffer, the cycling temperatures and the balance between the magnesium chloride and deoxynucleotide concentrations. Based on our experience, we propose a protocol for developing a multiplex PCR assay and suggest ways to overcome commonly encountered problems.
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Cited by
Point-of-Care Diagnostics for Global Health
Yager, P., Domingo, G. J., & Gerdes, J. (2008) · Annual Review of Biomedical Engineering · doi:10.1146/annurev.bioeng.10.061807.160524 · Cited by 1,076
Vankerckhoven, V., Van Autgaerden, T., Vael, C., Lammens, C., Chapelle, S., Rossi, R., Jabes, D., & Goossens, H. (2004) · Journal of Clinical Microbiology · doi:10.1128/jcm.42.10.4473-4479.2004 · Cited by 594
Multiplex PCR: Optimization and Application in Diagnostic Virology
Elnifro, E. M., Ashshi, A. M., Cooper, R. J., & Klapper, P. E. (2000) · Clinical Microbiology Reviews · doi:10.1128/cmr.13.4.559-570.2000 · Cited by 964
Predicting Stability of DNA Duplexes in Solutions Containing Magnesium and Monovalent Cations
Owczarzy, R., Moreira, B. G., You, Y., Behlke, M. A., & Walder, J. A. (2008) · Biochemistry · doi:10.1021/bi702363u · Cited by 363
Dubuc, C., Ruiz-Lambides, A., & Widdig, A. (2014) · Behavioral Ecology · doi:10.1093/beheco/aru052 · Cited by 76
Metadata
- Authors
- Henegariu, O., Heerema, N. A., Dlouhy, S. R., Vance, G. H., & Vogt, P. H.
- Venue
- BioTechniques
- Publisher
- Informa UK Limited
- Published
- 1997-09
- Volume
- 23
- Issue
- 3
- Pages
- 504-511
- ISSN
- 0736-6205 (ISSN-L)
- 1940-9818
- OpenAlex
- W2096298665
- Cited by
- 953
- References
- 0
- FWCI
- 4.28
- Type
- journal article
- Language
- en
- Open access
- gold
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- Henegariu, O., Heerema, N. A., Dlouhy, S. R., Vance, G. H., & Vogt, P. H. (1997). Multiplex PCR: Critical Parameters and Step-by-Step Protocol. BioTechniques. https://doi.org/10.2144/97233rr01
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.
