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Lecture
Lecture evaluations are here
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Laboratory
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Monday
9:00 to 12:30
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Proteomics and the two-hybrid system:
achievements and perspectives
Introduction to the yeast two-hybrid system
Yeast as a Genetic Tool
Two-hybrid system components
The interaction-trap system as an example
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12:30 to 13:30
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Break
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13:30 to 14:00
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Bait Assessment
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14:00 to 17:30
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Bait Assessment
Yeast transformation
Mating assay
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Tuesday
9:00 to 12:30
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Library Screening
Interaction mating
YTH-related techniques
False Positives
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12:30 to 13:30
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Break
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13:30 to 17:00
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Mating assay (continued)
Filter beta-gal assay
CHCl3 overlay beta-gal assay
Yeast DNA prep and PCR
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Wednesday
9:00 to 12:30
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Variations & New Developments
Dual Bait system
Split-ubiquitin system
other non- trancription-based systems
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12:30 to 13:30
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Break
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13:30 to 17:00
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Analyse PCR products
split-ubiquitin system,
LexA-Gal4 comparison:
replica-plating
Bait Assessment (results)
Yeast transformation (results)
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Thursday
9:00 to 12:00
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Two-hybrid: what are your chances on success?
Two-hybrid on the Internet
Specialized yeast two-hybrid systems
(one, 1.5, tri-hybrid, RNA etc.)
Bacterial two-hybrid systems
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12:30 to 13:30
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Break
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13:30 to 17:00
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Liquid beta-gal assay
Bacterial two-hybrid system:
transformation
Mating assay (results)
False positives: plating
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Friday
9:00 to 11:00
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Commercially available systems:
LexA- & Gal4-based systems
Bioinformatics: extending your clone in silico
Using YTH in pharmaceutical screening
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11:00 to 13:00
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Break
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split-ubiquitin system,
LexA-Gal4 comparison:
beta-gal assay by CHCl3 overlay
Bacterial two-hybrid system:
review plates
Results and Discussion
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14:00 to 17:00
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Questions and Answers; General Discussion
Screening done: what is next?
(Other techniques for detection of protein-protein interactions)
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