IGEM:PennState/Labbook/LucienWeiss

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 1. 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

1. 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]
2. Grown overnight at 37 degrees C on 50 micromolar Kan LB plates
3. 3mL cultures inoculated off of the plate and grown at 30 degrees C until OD590~.45
4. 3uL placed of receiver cells placed in various distances from 3uL inoculated sender colony.

Results:

Poster: Bacterial Relay Race .40% Eiken Agar with sender produced AHL tagged for degradation after 8 hours |thumb|center|.40% Eiken Agar with regular sender and receiver cells after 8 hours

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

March 13, 2007: Motility Assay

March 16, 2007: Motility Assay

March 29-April 1, 2007: Institute of Biological Engineering

Poster: Bacterial Relay Race IBE Poster

April 22, 2007: Project Proposal Presentations

Project Proposal: Pressure Sensing in Bacteria Slide1 Slide2 Slide3 Slide4 Slide5 Slide6 Slide7 Slide8

Project Proposal: Bi and Sept Stable Oscillator Slide1 Slide2 Slide3 Slide4 Slide5 Slide6 Slide7