Procedure for a high throughput RNAi screen for the second phase of the iBeetle screen

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In this proposal we describe in detail the screening procedure we developed for iBeetle, and the resources required at the two screening centers to conduct this genome-wide RNAi screen. As described in Part A Concept & Aims of this application, this procedure has been tested and optimized in a pilot screen. In this proposal we apply for the funds necessary to functionally analyze about half of the Tribolium genome: In the first grant period, we intend to test dsRNAs of 6,000 genes. To optimize the yield of the screen relative to the effort invested, each dsRNA will be injected into both, larvae and pupae. By this means, gene functions in a number of different processes during embryonic development as well as metamorphosis can be detected. Screening in two centers will ensure high efficiency by concentrating the practical expertise, will ensure uniform screening criteria, and will allow for division of labor and specialization in analysis/phenotype detection among the students, the TAs and the postdoc involved (the screeners )- Moreover, screening at two centers allows maximal use of the existing infrastructure.