High Throughput Methods in Proteomics

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David Wishart
University of Alberta
Edmonton, AB
david.wishart@ualberta.ca

Proteomics
Proteomics employs an incredibly diverse range of technologies including:
molecular biology
chromatography
electrophoresis
mass spectrometry
X-ray crystallography
NMR spectroscopy
microscopy
computational biology

Proteomics Tools
Molecular Biology Tools
Separation & Display Tools
Protein Identification Tools
Protein Structure Tools
Molecular Biology Tools
Northern/Southern Blotting
Differential Display
RNAi (small RNA interference)
Serial Analysis of Gene Expression (SAGE)
DNA Microarrays or Gene ChIP s
Yeast two-hybrid analysis
Immuno-precipitation/pull-down
GFP Tagging & Microscopy

SAGE
Principle is to convert every mRNA molecule into a short (10-14 base), unique tag. Equivalent to reducing all the people in a city into a telephone book with surnames
After creating the tags, these are assembled or concatenated into a long “list”
The list can be read using a DNA sequencer and the list compared to a database to ID genes or proteins and their frequency

SAGE Tools

SAGE of Yeast Chromosome