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Quantitative real‐time PCR for rapid and accurate titration of recombinant baculovirus particles
Author(s) -
Hitchman Richard B.,
Siaterli Evangelia A.,
Nixon Clare P.,
King Linda A.
Publication year - 2006
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.21177
Subject(s) - titration , recombinant dna , chromatography , real time polymerase chain reaction , chemistry , baculoviridae , virology , biology , biochemistry , gene , organic chemistry , spodoptera
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.