Abstract
Quantitative PCR-based methods have proven to be easy-to-use, cost-effective procedures for the quantification of viral gene expression and viral genome numbers. Quantitative PCR (qPCR) and quantitative reverse transcriptase-PCR (qRT-PCR) are rapid and sensitive approaches that can be used to pinpoint defects in viral DNA replication and transcriptional activity, respectively. Due to the significant nucleotide overlap between Poxviridae these methods can be employed across a wide range of viruses from this family. Here we provide methods for the quantification of vaccinia DNA replication by qPCR and quantification of the three classes of vaccinia gene transcription by qRT-PCR.
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Acknowledgments
HM and JM are supported by core funding to the MRC Laboratory for Molecular Cell Biology at University College London (J.M.), the European Research Council (649101-UbiProPox), and the UK Medical Research Council (MC_UU12018/7).
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Huttunen, M., Mercer, J. (2019). Quantitative PCR-Based Assessment of Vaccinia Virus RNA and DNA in Infected Cells. In: Mercer, J. (eds) Vaccinia Virus. Methods in Molecular Biology, vol 2023. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9593-6_12
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DOI: https://doi.org/10.1007/978-1-4939-9593-6_12
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-4939-9593-6
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