ARS | Publication request: Plasmid-Based Functional Proteomic Robotic Workcell Process for High-Throughput Screening of Multiplexed Libraries of Mutagenized Clones

Submitted to: Meeting Abstract
Publication Type: Abstract
Publication Acceptance Date: October 18, 2005
Publication Date: October 18, 2005
Citation: Hughes, S.R., Riedmuller, S., Mertens, J.A., Li, X., Qureshi, N., Bischoff, K.M., Jordan, D.B., Cotta, M.A., Farrelly, P. 2005. Plasmid-based functional proteomic robotic workcell process for high-throughput screening of multiplexed libraries of mutagenized clones [abstract]. Optimization high-throughput Cultures for Bioprocessing 2005. p. 3.

Technical Abstract: When screening large numbers of clones using an integrated plasmid-based functional proteomic robotic workcell, careful attention must be given to minimizing reagent costs. Much of the molecular biology for these processes uses Gateway system recombinational cloning. We have explored a picking strategy that "multiplexes" or combines several mutagenized clones in each well to reduce the amount of reagent required. The workcell system described makes possible fully automated multiplex picking routines followed by automated plasmid preparation and in vitro expression of proteins. The proteins having desired characteristics are then identified and isolated. Protocols are described for the entire process from colony picking to assay screening of "hit" wells.