✦ LIBER ✦

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

✍ Scribed by Richard B. Hitchman; Evangelia A. Siaterli; Clare P. Nixon; Linda A. King

Publisher: John Wiley and Sons
Year: 2007
Tongue: English
Weight: 120 KB
Volume: 96
Category: Article
ISSN: 0006-3592
DOI: 10.1002/bit.21177

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✦ Synopsis

Abstract

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 C~T~ 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.

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