Quantification of gene expression of Listeria monocytogenes by real-time reverse transcription PCR: optimization, evaluation and pitfalls
Werbrouck, H. Botteldoorn, N Uyttendaele, M. Herman, L. Van Coillie, E.
Werbrouck, H.
Botteldoorn, N
Uyttendaele, M.
Herman, L.
Van Coillie, E.
Citations
Altmetric:
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 (
Description
Date
2007-05-15
Journal Title
Journal ISSN
Volume Title
Publisher
Chapter title
Publication type
Peer reviewed scientific article
Collections
Research Projects
Organizational Units
Journal Issue
Keywords
a, ALL, Animals, article, AS, at, Belgium, biosynthesis, Cell, cells, chemistry, Comparative Study, data, Deoxyribonuclease I, Dna, DNA,Complementary, Efficiency, EVALUATION, expression, factors, Flemish, food, Food Microbiology, gene, Gene Expression, Gene Expression Profiling, general, genetics, growth, Humans, i, im, Institute, IS, isolation & purification, IT, journal, LEVEL, levels, Listeria, Listeria monocytogenes, metabolism, method, methods, need, needs, outcome, PCR, Print, Quantification, random, Research, Research Support, result, results, Reverse Transcriptase Polymerase Chain Reaction, Rna, RNA,Bacterial, SB - IM, SENSITIVITY, Sequence Analysis,DNA, standards, strain, study, technology, time, use