small molecule drugs that have either broad spectrum A high throughput filtration assay was developed antibacterial or antifungal activity. We used RNA polyand automated for screening compounds and natural merase isolated from Escherichia coli as our primary product extracts using 96-well filterplates. The selecbacterial high throughput target enzyme and contion of a hydrophobic membrane and appropriate capducted a promoter-specific transcription assay. For the ture conditions enabled us to perform the enzymatic fungal polymerase, we used Candida albicans RNA reaction, capture of products, vacuum filtration, and polymerase II as our target.
detection in a single filtration microplate, without any
To develop a high throughput screening assay for transfer step. The hydrophobic membrane has the adpolymerases, we needed to have an automated robotic vantage of preventing any loss of aqueous solution filtration assay. Filtration assays reported in literaduring the enzymatic reaction. The results are compature utilize either a one-plate system with simple vacrable with those obtained in a solid plate format foluum devices (3 -5) or a two-plate system with a cell lowed by transferring the reaction mixture to a tradiharvester (6, 7). In the two-plate system, one performs tional hydrophilic filterplate. This assay is being used the assay in a solid microplate, captures the reaction to screen for microbial RNA polymerase inhibitors as products, transfers the products to a filtration plate potential new drugs for treatment of emerging antibito separate free from bound reagents, and carries out otic-resistant bacterial and fungal infections. The fila detection step. Variations from well to well are gentration setup can be applied to a variety of assay syserally small if all the contents are transferred accutems.