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Quantification of gene expression of Listeria monocytogenes by real-time reverse transcription PCR: optimization,
Hadewig Werbrouck1, Nadine Botteldoorn, Mieke Uyttendaele
1Flemish Government, Institute for Agricultural and Fisheries Research, Unit Technology and Food, Brusselsesteenweg 370, 9090 Melle, Belgium. Hadewig.Werbrouck@ilvo.vlaanderen.be
Abstract:
In the current study, various steps in the real-time reverse transcription PCR (real-time RT-PCR) method for determination of RNA expression levels starting from different numbers of Listeria monocytogenes cells were evaluated and optimized. Our results showed that the RNA isolation method as well as the cDNA synthesis may influence the sensitivity of the procedure. For high bacterial cell numbers (10(9) bacterial cells), the RNAqueous kit and the RNeasy Mini kit were equally useful, whereas for low bacterial cell numbers (
