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Genome-scale RNAi screens for high-throughput phenotyping in bloodstream-form African trypanosomes.
- Source :
-
Nature protocols [Nat Protoc] 2015 Jan; Vol. 10 (1), pp. 106-33. Date of Electronic Publication: 2014 Dec 11. - Publication Year :
- 2015
-
Abstract
- The ability to simultaneously assess every gene in a genome for a role in a particular process has obvious appeal. This protocol describes how to perform genome-scale RNAi library screens in bloodstream-form African trypanosomes, a family of parasites that causes lethal human and animal diseases and also serves as a model for studies on basic aspects of eukaryotic biology and evolution. We discuss strain assembly, screen design and implementation, the RNAi target sequencing approach and hit validation, and we provide a step-by-step protocol. A screen can yield from one to thousands of 'hits' associated with the phenotype of interest. The screening protocol itself takes 2 weeks or less to be completed, and high-throughput sequencing may also be completed within weeks. Pre- and post-screen strain assembly, validation and follow-up can take several months, depending on the type of screen and the number of hits analyzed.
Details
- Language :
- English
- ISSN :
- 1750-2799
- Volume :
- 10
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Nature protocols
- Publication Type :
- Academic Journal
- Accession number :
- 25502887
- Full Text :
- https://doi.org/10.1038/nprot.2015.005