title: Rapid and cost-effective baculovirus sample preparation method as a viable alternative to conventional preparation for quantitative real-time PCR.
abstract: The increasing use of the baculovirus expression vector system (BEVS) has generated significant interest into techniques for quantifying baculovirus stocks. One method involves the use of quantitative real-time polymerase chain reaction (PCR). This study investigated simplifying baculovirus sample preparation for quantitative Real Time PCR to provide an alternative to current kit-based preparation methods. To achieve this goal, combinations of freeze/thaw cycles and Triton X-100 treatment were investigated. A treatment with only Triton X-100 was found to be sufficient to provide a simple, rapid and cheap alternative to kit-based preparation methods. This study also examined other factors such as primer choice to further examine the process of baculovirus quantitation by qPCR.
authors.affiliation: Department of Chemical Engineering, Waterloo Institute for Nanotechnology, University of Waterloo, Waterloo, ON, Canada N2L 3G1.
authors.forename: Steve
authors.initials: S
authors.lastname: George
authors.forename: Stanislav
authors.initials: S
authors.lastname: Sokolenko
authors.forename: Marc Gordon
authors.initials: MG
authors.lastname: Aucoin
mesh_terms: D016367:Baculoviridae / Q000235:genetics*; D008967:Molecular Biology / Q000191:economics / Q000379:methods*; D009696:Nucleic Acids / Q000235:genetics / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000191:economics / Q000379:methods*; D013048:Specimen Handling / Q000191:economics / Q000379:methods*; D013997:Time Factors
chemical_list: D009696:Nucleic Acids