Title: Use of gDNA as internal standard for gene expression in Staphylococci in vitro and in vivo
Authors: Vandecasteele, Stefaan ×
Peetermans, Willy
Merckx, Rita
Van Ranst, Marc
Van Eldere, Johan #
Issue Date: Mar-2002
Publisher: Academic Press
Series Title: Biochemical and Biophysical Research Communications vol:291 pages:528-534
Abstract: An internal RNA standard proved less suitable in bacterial gene expression experiments. We therefore developed a method for simultaneous RNA and gDNA (genomic DNA) isolation from in vitro and in vivo samples containing staphylococci and combined it with quantitative PCR. The reliability of gDNA for bacterial quantification and for standardisation in gene expression experiments was evaluated. Quantitative PCR proves equivalent to quantitative culture for in vitro samples, and superior for in vivo samples. In gene expression experiments, gDNA permits a good standardisation for the initial amount of bacteria. The average interassay variability of the in vitro expression is 20.1%. The in vivo intersample variability was 73.3%. This higher variability can be attributed to the biological variation of gene expression in vivo. This method permits exact quantification of the number of bacteria and the expression of genes in staphylococci in vivo (e.g., in biofilms, evolution in time) and in vitro.
ISSN: 0006-291X
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Laboratory of Clinical and Epidemiological Virology (Rega Institute)
Laboratory for Clinical Infectious and Inflammatory Disorders
Laboratory of Clinical Bacteriology and Mycology
Laboratory for Experimental and Clinical Microbiology (-)
× corresponding author
# (joint) last author

Files in This Item:

There are no files associated with this item.

Request a copy


All items in Lirias are protected by copyright, with all rights reserved.

© Web of science