The DOI matches its Crossref record; 38 of the 38 references it cites were found in a registry or an index.
qPCR primer design revisited
Open accessAbstract
Primers are arguably the single most critical components of any PCR assay, as their properties control the exquisite specificity and sensitivity that make this method uniquely powerful. Consequently, poor design combined with failure to optimise reaction conditions is likely to result in reduced technical precision and false positive or negative detection of amplification targets. Despite the framework provided by the MIQE guidelines and the accessibility of wide-ranging support from peer-reviewed publications, books and online sources as well as commercial companies, the design of many published assays continues to be less than optimal: primers often lack intended specificity, can form dimers, compete with template secondary structures at the primer binding sites or hybridise only within a narrow temperature range. We present an overview of the main steps in the primer design workflow, with data that illustrate some of the unexpected variability that often occurs when theory is translated into practice. We also strongly urge researchers to report as much information about their assays as possible in their publications.
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1 retracted
Lee, T. K., Murthy, S. R. K., Cawley, N. X., Dhanvantari, S., Hewitt, S. M., Lou, H., Lau, T. C. M., Ma, S., Huynh, T., WESLEY, R. A., et al. (2011) · Journal of Clinical Investigation · doi:10.1172/jci40433 · Cited by 62
RetractedThe MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments
Bustin, S. A., Benes, V., Garson, J. A., Hellemans, J., Huggett, J., Kubista, M., Mueller, R., Nolan, T., Pfaffl, M. W., Shipley, G. L., et al. (2009) · Clinical Chemistry · doi:10.1373/clinchem.2008.112797 · Cited by 16,303
Self-citationMfold web server for nucleic acid folding and hybridization prediction
Zuker, M. (2003) · Nucleic Acids Research · doi:10.1093/nar/gkg595 · Cited by 13,404
Epidemiologic classification of human papillomavirus types associated with cervical cancer
Muñoz, N., Bosch, F. X., de Sanjosé, S., Herrero, R., Castellsagué, X., Shah, K. V., Snijders, P. J. F., & Meijer, C. J. L. M. (2003) · New England Journal of Medicine · doi:10.1056/nejmoa021641 · Cited by 6,249
Quantification of mRNA using real-time RT-PCR
Nolan, T., Hands, R. E., & Bustin, S. A. (2006) · Nature Protocols · doi:10.1038/nprot.2006.236 · Cited by 2,247
Self-citationThe Thermodynamics of DNA Structural Motifs
SantaLucia, J., & Hicks, D. (2004) · Annual Review of Biophysics and Biomolecular Structure · doi:10.1146/annurev.biophys.32.110601.141800 · Cited by 1,314
Huggett, J. F., Foy, C. A., Benes, V., Emslie, K., Garson, J. A., Haynes, R., Hellemans, J., Kubista, M., Mueller, R. D., Nolan, T., et al. (2013) · Clinical Chemistry · doi:10.1373/clinchem.2013.206375 · Cited by 859
Self-citationZipper, H. (2004) · Nucleic Acids Research · doi:10.1093/nar/gnh101 · Cited by 791
Bustin, S. A., Beaulieu, J.-F., Huggett, J., Jaggi, R., Kibenge, F. S., Olsvik, P. A., Penning, L. C., & Toegel, S. (2010) · BMC Molecular Biology · doi:10.1186/1471-2199-11-74 · Cited by 700
Self-citationForootan, A., Sjöback, R., Björkman, J., Sjögreen, B., Linz, L., & Kubista, M. (2017) · Biomolecular Detection and Quantification · doi:10.1016/j.bdq.2017.04.001 · Cited by 578
Cited by
Salmon, M., White, H. E., Zizkova, H., Gottschalk, A., Motlova, E., Cerveira, N., Colomer, D., Coriu, D., Franke, G. N., Gottardi, E., et al. (2022) · Leukemia · doi:10.1038/s41375-022-01612-2 · Cited by 22
Metadata
- Authors
- Bustin, S., & Huggett, J.
- Venue
- Biomolecular Detection and Quantification
- Publisher
- Elsevier BV
- Published
- 2017-12
- Volume
- 14
- Pages
- 19-28
- ISSN
- 2214-7535 (ISSN-L)
- OpenAlex
- W2768784295
- Cited by
- 312
- References
- 38
- FWCI
- 8.74
- Type
- journal article
- Language
- en
- Open access
- hybrid
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