Large-scale Gene Knockdown in C. elegans Using dsRNA Feeding Libraries to Generate Robust Loss-of-function Phenotypes

Kathryn N. Maher; Mary Catanese; Daniel L. Chase

doi:10.3791/50693

/

Storskala Gene Knockdown i C. elegans Bruke dsrna Fôrings biblioteker for å generere en solid tap-av-funksjon Fenotyper

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Large-scale Gene Knockdown in C. elegans Using dsRNA Feeding Libraries to Generate Robust Loss-of-function Phenotypes

Storskala Gene Knockdown i C. elegans Bruke dsrna Fôrings biblioteker for å generere en solid tap-av-funksjon Fenotyper

DOI:

10.3791/50693-v

•

18:38 min

•

September 25, 2013

•

Kathryn N. Maher, Mary Catanese, Daniel L. Chase

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

Mens dsrna fôring i C. elegans er et kraftig verktøy for å vurdere gen-funksjon, gjeldende protokoller for storskala fôrings skjermene resultere i variable knockdown effektivitet. Vi beskriver en forbedret protokoll for å utføre storskala RNAi fôring skjermer som resulterer i svært effektiv og reproduserbar knockdown av genuttrykk.

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