SEED/2009/Day 8

Morning

 * Discuss beta-gal assay and AHL-inducible lacZ
 * Take 20ul of the culture into 80ul of the permeabilization solution (directly in cuvettes)
 * Leave at 30C over lunch
 * Measure the OD600 of the original cultures while waiting

Afternoon

 * Add 600ul substrate solution to permeabilized cells
 * Every 20 minutes, make another measurement and record time

Homework

 * Analyze data
 * Plot results
 * Compare results with existing F2620 data

Instructor Pre-prep

 * grow up 1ml cultures expressing F2620.lacZa (BBa_T9003) with IPTG and different amounts of AHL
 * 0, 1E-5, 1E-6, 1E-7, 1E-8, 1E-9, 1E-10, 1E-11
 * grow up 1ml control cultures of cells with and w/o lacZa expression plasmid
 * Make solutions as specified in Beta-Galactosidase_Assay_(A_better_Miller)