Abstract
Hyperbranching Rolling Circle Amplification (HRCA) is a technique derived from Rolling Circle Amplification (RCA) in which DNA polymerase replicates circularized oligonucleotide probes under isothermal conditions with either linear or geometric kinetics. Since its first introduction HRCA has been proven to be a robust DNA amplification technique for detecting pathogenic fungi and other applications, allowing rapid detection of nucleic-acid sequences with high specificity. Here we describe an improved protocol of HRCA, which is both specific and sensitive for detecting low copy numbers of template DNA. The test can be performed within one working day in routine molecular labs.
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Acknowledgments
This work was supported by The Federation of European Biochemical Societies (FEBS) Chinese European Visiting Fellowships (CEVF-00007), and was partly supported by the International Program of Project 985 at Sun Yat-Sen University.
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Sun, J., de Hoog, S. (2013). Hyperbranching Rolling Circle Amplification, An Improved Protocol for Discriminating Between Closely Related Fungal Species. In: O'Connor, L., Glynn, B. (eds) Fungal Diagnostics. Methods in Molecular Biology, vol 968. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-257-5_13
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DOI: https://doi.org/10.1007/978-1-62703-257-5_13
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Publisher Name: Humana Press, Totowa, NJ
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