Abstract
ProbeLibrary, a fast, specific and flexible format for quantitative real-time PCR, is described. The ProbeLibrary concept is based on the fact that just 90 short probes provide transcriptome-wide coverage in most organisms. These short probes are highly specific and possess the high melting temperature (Tm) required for real-time PCR owing to the incorporation of locked nucleic acid (LNA). The probes are dual-labeled fluorogenic probes that are used in standard real-time PCR protocols, with standard instrumentation. ProbeLibrary probes are used in assays designed using free, web-based assay design software, termed ProbeFinder. In seconds, ProbeFinder designs highly specific intron-spanning assays for a target transcript. The system allows multiple design options, including designs for transcript variants and gene family members. Assay design success rate is 96%. This combination of extremely fast assay design and instant availability of the probes leads to faster implementation of real-time PCR assays.
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References
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Acknowledgements
Technical assistance from S. Ludvigsen, M.B. Mogensen and M. Bjørn is gratefully acknowledged.
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This article was submitted to Nature Methods by a commercial organization and has not been peer reviewed. Nature Methods takes no responsibility for the accuracy or otherwise of the information provided.
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Mouritzen, P., Noerholm, M., Nielsen, P. et al. ProbeLibrary: A new method for faster design and execution of quantitative real-time PCR. Nat Methods 2, 313–316 (2005). https://doi.org/10.1038/nmeth0405-313
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DOI: https://doi.org/10.1038/nmeth0405-313
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