IGEM:IMPERIAL/2008/Prototype/Wetlab/Protocols

= Motility Experiments =

Isolating motile derivatives

 * 1) Streak each strain carefully at one point of a 0.7% TBAB agar plate.
 * 2) Incubate overnight at 37ºC without inverting the plates.
 * 3) Motile cells will swarm over the surface of the plate and should be picked from as far from the site of inoculation as possible. If the strain was non-motile there will be a contained streak with a few dendritic swarms emerging from the streak. Pick from the top of one of these swarms to recover a motile derivative.

Microscope Protocol
(For GFP Microscopy)
 * 1) Grow cells overnight at 22°C in LB to 1 OD_600
 * 2) Pellet 1ml and resuspend in 0.1ml PBS
 * 3) Stain membranes by resuspension in 50μL of PBS containing 5μg/mL FM4-64 and incubate for 10min at room temperature.
 * 4) Observe samples by spotting 3μL of suspension on a glass microscope slide and immobilise with poly-L-lysine-treated glass coverslip.
 * 5) Capture images