Quantification of gene expression
Gene expression analysis is fundamental in many fields of research and allows to understand dynamic changes in a bacterium, cell, tissue or organism. Indeed, if the genome is identicial in each cells of a an organism, genes can be expressed in a specific and different manners, depending on time (specific to development stage), on space (expression specific to a cell type or tissue) and/or on characteristics of a given state (normal, pathological, in response to stimuli, ...).
SMALTIS proposes to specifically study the expression of your genes by quantifying the number or your targetted mRNA thanks to its real-time PCR platform.
Depending on your needs, our staff will carry out your project following several steps:
STEP 1: Feasability Study
This step includes a litterature review to analyse already publised data allowing our staff to:
- Identify existing commercial quantification kits
- Design specific primers and probes (SybrGreen or TaqMan technologies)
- Develop a qPCR protocol
- Determine the optimal amplification conditions
- Verify the specificity of primers and probes
- Validate the choice of housekeeping genes
During this step, the analysis of a limited number os samples will also be done to validate the method. A detailed and illustrated report will be written to summarise the data.
STEP 2: Routine Analysis
This step allow us to analyse all your samples according to the previously validated method
Our platform equipped with a RotorGene 6000 and CFX Connect allows us to process a few samples at several hundred simultaneously.
Examples of achivements:
- Expression analysis of virulence and resistance genes in strains of Klebsiella pneumoniae
- Study of the inductive effect of antibiotics on the expression of efflux systems in Pseudomonas aeruginosa
- Detection of episomal DNA in blood samples collected using PAXGene® Technology
- Gene expression analysis of genes encoding IL-6, IL-8 and TNFα cytokines in cells previously infected with different bacterial strains
- Quantification of several genes in transfected cell lines.
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PCR & Sequencing
Rolling Circle Amplification