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在大规模的基因敲除<em> C。线虫</em>使用双链RNA料库产生失去功能的鲁棒表型
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JoVE Journal Biology
Large-scale Gene Knockdown in C. elegans Using dsRNA Feeding Libraries to Generate Robust Loss-of-function Phenotypes
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在大规模的基因敲除<em> C。线虫</em>使用双链RNA料库产生失去功能的鲁棒表型

DOI:
10.3791/50693-v

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18:38 min

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September 25, 2013

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, ,
1Molecular and Cellular Biology Program,University of Massachusetts, Amherst, 2Neuroscience and Behavior Program,University of Massachusetts, Amherst, 3Department of Biochemistry and Molecular Biology,University of Massachusetts, Amherst

Chapters

  • 00:05Title
  • 02:54How to Determine Plasmid Loss
  • 04:32Making Temporary Copies of the Library Omniplates
  • 06:35Preparation of Bacteria for Feeding Worms
  • 09:28Determining the Concentration of L1 Arrested C. elegans Larvae
  • 10:53dsRNA Feeding Screen
  • 12:37Results: Effects of egl-30, dpy-17, pat-10, and unc-4 Knockdown
  • 17:50Conclusion

Summary

Automatic Translation

Automatic Translation

而喂养的dsRNA在C线虫是一种功能强大的工具来评估基因功能，为大规模饲养屏幕当前的协议导致变量击倒效率。我们描述了一种改进的协议用于进行大规模的RNAi馈送屏幕，导致基因表达的高度有效的和可再现的击倒。

Tags

RNA InterferenceRNAiDsRNA FeedingC. ElegansGene KnockdownLoss-of-function PhenotypesHigh-throughput ScreeningPlasmid LossProtocolNeurons

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