IGEM:University of Chicago/2009/Notebook/Paraoxon Biosensor/2009/07/30

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July 30th, 2009
Fluorescence and Viability Assay

Take overnight cells, dilute to 0.05, and have both a non-PNP control and a PNP at 1 mM test. Do this for both 4.1 and 4.2 strains.

Below is a chart for the assay results. Induced with PNP at 1:48 p.m. Might see a decline in growth rate. Maybe glowing, also. 1mM

4.1 cont OD 1:30 = 0.152, OD 3:00 = 0.242, OD 4:30 = 0.438, OD 6:00 = 0.641

4.1 PNP OD 1:30 = 0.148, OD 3:00 = 0.210, OD 4:30 = 0.260, OD 6:00 = 0.355

4.2 cont OD 1:30 = 0.284, OD 3:00 = 0.209, OD 4:30 = 0.465, OD 6:00 = 0.635

4.2 PNP OD 1:30 = 0.131, OD 3:00 = 0.273, OD 4:30 = 0.224, OD 6:00 = 0.419

Trans 4 OD 1:30 = 0.472

Trans 4 OD 1:30 = 0.421

Transformation 4 (longtine PCR fragments)

- Attempt to transform Longtine PCR fragments into SKY816. OD average of two (10 ml) cultures was approximately 0.44. Hopefully, all will transform. If not all, hopefully fragments 1 and 5.

-PEG mixture seems a bit viscous. Not sure if it's correct or not....


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