Applications of Microarrays
09.30 - 17.00 14 May 2007
This course will be held prior to the Advances in Microarray Technology Conference.
Who should attend?
The course will be suitable for scientists, technicians and engineers. It is helpful to be familiar with genes, RNA and proteins to follow the course. The course is for people that consider working with microarrays and for those who are currently working with some applications of microarrays but want a broader view on the subject.
Learning Objectives
1. Master the basics of microarray including surface chemistries, fabrication,
hybridisation and detection technologies.
2. Understanding DNA microarray probe design for SNP detection and bacterial
diagnostics.
3. Fundamentals of gene expression analysis using DNA microarrays
4. Protein microarrays, fabrication and applications
5. Quantification of drugs using microarrays
Topics and Course Organisation
Fabrication and using microarrays
1. Immobilisation of DNA to different surfaces
a. Commercial slides and do it yourself slides
b. Surface chemistries
c. Probe modifications
2. Fabrication of microarrays
a. Contact printing
b. Non-contact printing
c. In situ synthesis of DNA
3. Hybridisation
a. Diffusion based
b. Mixing based
4. Detection
a. Fluorescence
b. Luminescence
c. Colorimetric
Demonstration of the effects of stringency in SNP genotyping by allele specific genotyping
1. Possible application of DNA microarrays: linkage studies and
diagnostics
2. Designing a microarray for bacterial diagnostics
3. Basics of allele specific hybridisation
4. Designing a DNA microarrays for SNP/small mutation detection as
exemplified by the globin gene
5. Minisequencing/primer extension on arrays
Genexpression studies using DNA microarrays
1. Examples of applications
a. Basic research on cell physiology
b. Cancer classifier
2. Platforms
a. Home made arrays/cDNA arrays
b. Affymetrix
c. Agilent
3. Production of target
a. Types of RNA (total RNA vs mRNA), quality control issues
b. Priming methods
c. Reverse transcription
d. T7 amplification
4. Hybridisation formats; co-hybridisation and parallel hybridisation
5. Detection
6. Bioinformatics
7. MIAME and importance of repeating experiments
Protein microarrays
1. Comparison between DNA and protein microarrays
2. Basics
a. Fabrication issues, orientation of protein on the surface and
keeping protein active after printing
b. Target preparation/target labelling
2. Applications
a. Immunoarrays
b. Protein arrays
i. Diagnostics (allergen microarrays)
ii. Proteome arrays
c. Small molecule arrays
Other types of arrays
1. Comparative genomic hybridisation
2. Genexpression studies on few cells
3. Global promoter studies using microarrays
a. Chromatin Immunoprecipitation using DNA miroarrays
b. Double stranded DNA microarrays
4. Transfecting cells on microarrays
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