To begin, transfer the hydrogels to 1.5 milliliter micro centrifuge tubes, trimming the gels with a scalpel if necessary to make them fit in the wells. In another 1.5 milliliter micro centrifuge tube, combine 10 microliters of the cell-free extract with 25 microliters of two x cell-free protein synthesis buffer and four micrograms of plasmid DNA. Make up to a total volume of 50 microliters with double distilled water to prepare the cell-free protein synthesis solution.
Pipette the solution onto the freeze dried hydrogels, and allow the gels to soak in the cell-free protein synthesis system for five to 10 minutes at room temperature. Transfer the gels to a black 384 well microtiter plate using a spatula. Then, transfer the microtiter plate to a plate reader, and use the plate reader settings shown on the screen for fluorescence detection and analysis.
