Quantitative real-time PCR for rapid and accurate titration of recombinant baculovirus particles

We describe the use of quantitative PCR (QPCR) to titer recombinant baculoviruses. Custom primers and probe were designed to gp64 and used to calculate a standard curve of QPCR derived titers from dilutions of a previously titrated baculovirus stock. Each dilution was titrated by both plaque assay a...

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Bibliographic Details
Published in:Biotechnology and bioengineering Vol. 96; no. 4; pp. 810 - 814
Main Authors: Hitchman, Richard B., Siaterli, Evangelia A., Nixon, Clare P., King, Linda A.
Format: Journal Article
Language:English
Published: Hoboken Wiley Subscription Services, Inc., A Wiley Company 01-03-2007
Wiley
Wiley Subscription Services, Inc
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Summary:We describe the use of quantitative PCR (QPCR) to titer recombinant baculoviruses. Custom primers and probe were designed to gp64 and used to calculate a standard curve of QPCR derived titers from dilutions of a previously titrated baculovirus stock. Each dilution was titrated by both plaque assay and QPCR, producing a consistent and reproducible inverse relationship between CT and plaque forming units per milliliter. No significant difference was observed between titers produced by QPCR and plaque assay for 12 recombinant viruses, confirming the validity of this technique as a rapid and accurate method of baculovirus titration. Biotechnol. Bioeng. 2007;96:810–814. © 2006 Wiley Periodicals, Inc.
Bibliography:South East Proof of Concept funding (SEPOC)
The Biotechnology and Biological Sciences Research Council (BBSRC)
istex:83FF4217EF9B64294BD8D55362FFB0C30690E02E
ArticleID:BIT21177
ark:/67375/WNG-BS7KZNQW-6
Richard B. Hitchman and Evangelia A. Siaterli contributed equally to this work.
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ISSN:0006-3592
1097-0290
DOI:10.1002/bit.21177