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PMID: 15644175 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

High-throughput RNA interference screens in Drosophila tissue culture cells.

Methods in enzymology ·Vol. 392 ·2005-00-00 ·Pages 55-73

Armknecht S, Boutros M, Kiger A, Nybakken K, Mathey-Prevot B, Perrimon N

Abstract

This chapter describes the method used to conduct high-throughput screening (HTs) by RNA interference in Drosophila tissue culture cells. It covers four main topics: (1) a brief description of the existing platforms to conduct RNAi-screens in cell-based assays; (2) a table of the Drosophila cell lines available for these screens and a brief mention of the need to establish other cell lines as well as cultures of primary cells; (3) a discussion of the considerations and protocols involved in establishing assays suitable for HTS in a 384-well format; and (A) a summary of the various ways of handling raw data from an ongoing screen, with special emphasis on how to apply normalization for experimental variation and statistical filters to sort out noise from signals.

MeSH Terms
Animals Cell Line Drosophila/cytology,genetics Genes, Reporter RNA Interference Tissue Culture Techniques Transfection
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Armknecht Susan
Department of Genetics, Harvard Medical School, Drosophila RNAi Screening Center, Boston, MA 02115, USA.
Boutros Michael
Kiger Amy
Nybakken Kent
Mathey-Prevot Bernard
Perrimon Norbert
Article Info
Journal
Methods in enzymology
Abbr.
Methods Enzymol
ISSN
0076-6879
Published
2005-00-00
Pages
55-73
Language
English
Region
United States
NLM ID
0212271
Subset
IM
Grants
NIGMS NIH HHS · R01 GM078176 · United States
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