Rapid and cost-effective baculovirus sample preparation method as a viable alternative to conventional preparation for quantitative real-time PCR

J Virol Methods. 2012 Jun;182(1-2):27-36. doi: 10.1016/j.jviromet.2012.03.001. Epub 2012 Mar 8.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Baculoviridae / genetics*
  • Molecular Biology / economics
  • Molecular Biology / methods*
  • Nucleic Acids / genetics
  • Nucleic Acids / isolation & purification*
  • Real-Time Polymerase Chain Reaction / economics
  • Real-Time Polymerase Chain Reaction / methods*
  • Specimen Handling / economics
  • Specimen Handling / methods*
  • Time Factors

Substances

  • Nucleic Acids