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Quantitative and Automated High-throughput Genome-wide RNAi Screens in C. elegans
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Quantitative and Automated High-throughput Genome-wide RNAi Screens in C. elegans

Quantitative and Automated High-throughput Genome-wide RNAi Screens in C. elegans

DOI:
10.3791/3448-v

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10:58 min

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February 27, 2012

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, , ,
1Centre d’Immunologie de Marseille-Luminy,Université de la Méditerranée

Chapters

  • 00:05Title
  • 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

Summary

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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.

Tags

QuantitativeAutomatedHigh-throughputGenome-wideRNAi ScreensC. ElegansGene FunctionKnockdownBacteriaDsRNALibrariesFunctional Genomic StudiesLaborious MethodsSemi-automated ProtocolsMicroscopic ImagingImage AnalysisRobotic HandlingCOPAS BiosortMBioLIMSData ManagementSample TrackingSolid Medium PlatesHost-pathogen InteractionsAnti-fungal Innate Immunity

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