Gene Expression
November 10
Papers to discuss:
Vasemagi and Primmer 2005.pdf
Challanges for identifying funtionally important genetic variation: the promise of combining complementary research strategies
Molecular Ecology (2005) 14, 3623-3642
michel et al 2004.pdf
Somatic mutation-mediated evolution of herbicide resistance in the nonindigenous invasive plant hydrilla (Hydrilla verticillata) by Michel et al. Molecular Ecology 13:3229-3237.
Hinfray et al.pdf
Brain and Gonadal Aromatase as Potential Targets of Endocrine Disrupting Chemicals in a Model Species, the Zebrafish (Danio rerio)
Sequence polymorphism can produce serious artefacts in real-time PCR assays: hard lessons from Pacific oysters.
BMC Genomics. 2008 May 20;9:234.
Knudsen et al.pdf
Evaluation of the Branched-Chain DNA Assay for Measurement of RNA in Formalin-Fixed Tissues
Journal of Molecular Diagnostics, Vol.10, No.2, March 2008
Website about use of zebrafish as a research model.
Technology for creating zebrafish knockouts
Learn more about endocrine disruptors
![Estrogen Receptor Binding Estrogen Receptor Binding](http://e.hormone.tulane.edu/learning/images/estrogen-docking.gif) |
Estrogen Receptor Binding |
QuantiGene® Reagent System----
QuantiGene® Reagent System
QuantiGene assays bring the power of branched DNA (bDNA) technology to basic and pre-clinical research. The bDNA assay is a sandwich nucleic acid hybridization method that uses bDNA molecules to amplify signal from captured target RNA. bDNA technology forms the basis of the FDA-approved clinical diagnostic VERSANT 3.0 assays for HIV, HCV and HBV viral load, that are offered commercially by Siemens and have been in use for over a decade. Another advantage of bDNA assays is that RNA is measured directly from the sample source, without RNA purification or enzymatic manipulation, thereby avoiding inefficiencies and variability introduced by or errors inherent to these processes.
Principle of QuantiGene Assay
Principle of QuantiGene Assay
Step 1
Lyse cells to release mRNA in the presence of target probes.
Target mRNA from lysed cells is captured by hybridization and
transferred to the Capture Plate.
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![external image qg_assay1.gif external image qg_assay1.gif](http://www.panomics.com/images/qg_assay1.gif)
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Step 2
Signal amplification is performed by hybridization of the
bDNA Amplifier and Label Probe.
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![external image qg_assay2.gif external image qg_assay2.gif](http://www.panomics.com/images/qg_assay2.gif)
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Step 3
Addition of chemiluminescence substrate yields a luminescent
signal that is proportional to the amount of mRNA present in
the sample.
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![external image qg_assay3.gif external image qg_assay3.gif](http://www.panomics.com/images/qg_assay3.gif)
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Hydrilla in Washington
Click on the presentation below for hydrilla info in Florida
Schardt_%20Jeff.pdf
Other approaches for characterizing gene expression:
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Additional Papers on the topic:
Canales et al.pdf
Evaluation of DNA microarray results with quantitative gene expression platforms
Nature Biotechnology Vol. 24, No. 9, September 2006
Trant et al.pdf
Developmental expression of cytochrome P450 aromatase genes (CYP19a and CYP19b) in zebrafish fry (Danio rerio)
Journal of Experimental Zoology 290:475-483.
EHP114pa700PDF.PDF
An easy read about biomarkers and environmental health