| Solution Information | help | |
| Enzyme: | Protein RecA | |
| inhibitor: | BDBM70786 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Positive control drugs are clavulanic acid (beta-lactamase inhibitor) and Amikacin (both at 2.5ug/mL). All test compounds are screened at 25uM concentration. Compounds and controls are added to the plates at 2X concentration and given to the BSL3 personnel in charge of the assay execution. Assay in 384 well plate format contains 25uL of media and drugs, 25uL of bug and 9uL of Alamar Blue. 1. Day -5: M. tuberculosis culture: In the BSL3 laboratory, a culture of M. tuberculosis H37Rv is started from frozen stock in 10ml 7H9 medium without Tween-80 (1:20 dilution of culture; 500ul stock to 9.5ml medium). Grow M. tuberculosis culture at 37C for five days in a shaking water bath, until OD600 > 0.3, <0.9. 2. Day +1: Assay setup: Initial setup of plates occurs outside the BSL3 lab. The Biomek FX is used to add compounds, controls, and media to Corning 384-well black clear bottom plates (catalog no.3712) (25uL/well). Plates are drugged in duplicate, for screening in the presence and abse | |
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