Fig. 3.

High-throughput analytical scale protein expression screening using robotic methods. This schematic shows the step-by-step procedure used for small-scale expression screening. Completely automated steps are shown in blue, and partially automated steps are shown in red. Briefly, initial cultures are grown in 2.2 ml 96-well S-Blocks (Qiagen), followed by subculturing in 24 Well Block (Qiagen). Following overnight incubation the cultures are transferred into two separate S-Blocks (1 ml per respective well) and harvested by centrifugation (3000 × g, 10 minutes). The media is discarded and the cell pellet is resuspended in 100 μl lysis buffer and transferred to a 96-Well Round Bottom plate (Greiner). Following sonication a 30 μl aliquot of the total cellular lysate (Tot) is transferred to a new plate. The remainder is centrifuged for 10 minutes at 3000 × g, and a 30 μl aliquot of the supernatant (Sol) is transferred to a new plate. Equal amounts of Sol and Tot are added to adjacent wells for SDS-PAGE analysis.