Article ID Journal Published Year Pages File Type
3406817 Journal of Virological Methods 2012 10 Pages PDF
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.

► Comparison of qPCR quantitation of baculovirus using sequences from the Ie-1 and Gp-64 genes. ► Baculovirus DNA preparation using Triton X-100. ► Comparison of baculovirus titers using qPCR and flow cytometry.

Related Topics
Life Sciences Immunology and Microbiology Virology
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