Design and implementation of high-throughput RNAi screens in cultured Drosophila cells.

Published

Journal Article

This protocol describes the various steps and considerations involved in planning and carrying out RNA interference (RNAi) genome-wide screens in cultured Drosophila cells. We focus largely on the procedures that have been modified as a result of our experience over the past 3 years and of our better understanding of the underlying technology. Specifically, our protocol offers a set of suggestions and considerations for screen optimization and a step-by-step description of the procedures successfully used at the Drosophila RNAi Screening Center for screen implementation, data collection and analysis to identify potential hits. In addition, this protocol briefly covers postscreen analysis approaches that are often needed to finalize the hit list. Depending on the scope of the screen and subsequent analysis and validation involved, the full protocol can take anywhere from 3 months to 2 years to complete.

Full Text

Duke Authors

Cited Authors

  • Ramadan, N; Flockhart, I; Booker, M; Perrimon, N; Mathey-Prevot, B

Published Date

  • 2007

Published In

Volume / Issue

  • 2 / 9

Start / End Page

  • 2245 - 2264

PubMed ID

  • 17853882

Pubmed Central ID

  • 17853882

Electronic International Standard Serial Number (EISSN)

  • 1750-2799

Digital Object Identifier (DOI)

  • 10.1038/nprot.2007.250

Language

  • eng

Conference Location

  • England