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RNA Interference (RNAi) by Bacterial Feeding

Fanglian He Fanglian He

Published: Apr 20, 2011 DOI: 10.21769/BioProtoc.59 Views: 27605

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Abstract

There are 3 ways to perform RNAi in worms: microinjection, soaking and feeding. In the feeding protocol, RNAi is induced by cultivating worms on bacteria expressing gene-specific dsRNA. dsRNA is expressed in E. coli and ingested by worms. This protocol describes the feeding protocol to induce RNAi.

Materials and Reagents

  1. C. elegans RNAi clone/libraries (Open Biosystems or Source BioScience LifeSciences)
  2. Ampicillin (IBI Scientific)
  3. Tetracycline
  4. IPTG (Gold Biotechnology)
  5. LB agar medium: Agar (BD Biosciences), tryptone (BD Biosciences), yeast extract (BD Biosciences), NaCl (Research Organics)
  6. RNAi plate (NGM/IPTG/Ampicillin) (see Recipes)

Equipment

  1. Aluminum foil
  2. Petri dish (35 x 10 mm)

Procedure

Copyright: © 2011 The Authors; exclusive licensee Bio-protocol LLC.

Category

Molecular Biology > RNA > RNA interference

Molecular Biology > RNA > Transfection

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