IGEM:IMPERIAL/2006/Protocols/I13207

I13207 AiiA Test Construct Protocol

 * Culture bacteria (DH5a) in LB medium, incubate overnight
 * Transfer plasmids with desired part via electroporation
 * Incubate overnight
 * Transfer colonies into liquid medium (amp resistant, 2ml)
 * Wait until optical density is 0.1


 * Make 20mL of 1% arabinose LB solution
 * Then make the following solutions:


 * Transfer arabinose solutions into 5 different labeled tubes
 * One well is filled with pure LB media in all experiments as a control
 * Transfer 2 mL of each arabinose solution into separate clear tubes
 * Add 200ul of DH5a culture into the tubes
 * Assay with an automatically repeating protocol of absorbance measurements (600nm filter, 0.1 second absorbance through approximately 0.5cm of fluid)
 * Fluorescence readings (488nm excitation filter, 525nm emission filter, 0.5 second, CW lamp energy 12902 units)
 * Shake (1 mm, linear, normal speed)
 * Time between measurements should be 20 minutes
 * Time period for measurements should be ~5 h


 * Perform 2 tests: one with 0.5ml of 0.5M ZnCl2 in the LB medium and one without