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Quantitative and Automated High-throughput Genome-wide RNAi Screens in C. elegans
Bölümler
- 00:05Başlık
- 01:33Day 1, Preparation of 96-well NGM RNAi Plates, 96-DeppWell LB Plates, and RNAi Bacterial Clone Culturing
- 03:14Day 2, Seeding RNAi Bacterial Clones onto 96-well NGM RNAi Plates
- 04:46Days 3 and 4, RNAi Feeding of Worms and Infection of Worms with D. coniospora Spores
- 06:02Day 5, Observation and Storage of Plates Prior to Automated Quantitative Analysis
- 07:29Representative RNAi Results
- 10:27Conclusion
We describe a protocol using C. elegans and RNAi feeding libraries that allows automated measurement of multiple parameters such as fluorescence, size and opacity of individual worms in a population. We give one example of a screen to identify genes involved in anti-fungal innate immunity in C. elegans.
Etiketler
QuantitativeAutomatedHigh-throughputGenome-wideRNAi ScreensC. ElegansGene FunctionKnockdownBacteriaDsRNALibrariesFunctional Genomic StudiesLaborious MethodsSemi-automated ProtocolsMicroscopic ImagingImage AnalysisRobotic HandlingCOPAS BiosortMBioLIMSData ManagementSample TrackingSolid Medium PlatesHost-pathogen InteractionsAnti-fungal Innate Immunity
