Lucien Weiss Lab Notebook

Penn State iGEM 2005-2007
- [iGEM 2005]
- [iGEM 2006]
- [iGEM 2007]
Penn State Undergraduate
Email luw134@psu.edu
Phone (814) 880 5909

March 12, 2007: Motility Assay

Experiment Aims

Test motility on a variety of surface hardnesses in order to determine optimal level to use for assays.

Procedure

Eiken Agar Plates were made using a modified version of general procedure IGEM:PennState/2006/Eikenplates. Increased agar concentrations were made to increase hardness of gel and reduce the speed of motility. .20% Eiken Agar Plates:

.25g Bactotryptone
.20g NaCl
.15g sodium citrate
.05g Eiken Agar (for 20EikenAgar)

.30% Eiken Agar Plates:

.25g Bactotryptone
.20g NaCl
.15g sodium citrate
.075g Eiken Agar (for 30EikenAgar)

.40% Eiken Agar Plates:

.25g Bactotryptone
.20g NaCl
.15g sodium citrate
.10g Eiken Agar (for 40EikenAgar

Constructs were transformed into RP3087

YFP Sender Cells-Sender (LVA+)-RP3087 w/ [I14032+B0030+C0161 A3]+E0430
Sender Cells-Sender-RP3087 w/ I14032+B0030+C0161 (B2)
Receiver-RFP+Receiver-RP3087 w/ I13321+[J09855+B0033+J09274]

Grown overnight at 37 degrees C on 50 micromolar Kan LB plates
3mL cultures inoculated off of the plate and grown at 30 degrees C until OD590~.45
3uL placed of receiver cells placed in various distances from 3uL inoculated sender colony.

Results:

Observations: .20% agar results in good motility but is very fragile .30% agar results in fair motility and is much easier to handle .40% agar results in no motility

Conclusions:

Future Experiments: Retry .20% Agar experiments with set distances away from sender Experiment with temperature changes