IGEM:University of Chicago/2009/Notebook/Paraoxon Biosensor/2009/07/21

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July 21st, 2009

Growth assay using delta35 cultures 5.

Set up: dilute 1:100 in YPD and YPD + 0.5 mM paraoxon. Assay used by schofield, actual [] is 4.6

5.5 ml cultures

- (+) paraoxon

            - ul 20 mM paraoxon = 125 ul
            - ul YPD 4875 ul
            - culture 500 ul
            - total = 5.5 ml


- (-) paraoxon

            - ul 20 mM paraoxon = 0 ul
            - ul YPD 5000 ul
            - culture 500 ul
            - total = 5 ml

Start at 10:35. Take at least 6 time points

OD time points


- (+) paraoxon

            - 10:35 = 0.014
            - 12:05 = 0.030
            - 1:45 = 0.043
            - 4:05 = 0.207
            - 6:00 = 0.546

July 22nd reading : 10:00 a.m. = 1.559

- (-) paraoxon

            - 10:35 = 0.012
            - 12:05 = 0.030
            - 1:45 = 0.040
            - 4:05 = 0.215
            - 6:00 = 0.536

July 22nd reading : 10:00 a.m. = 1.554

Should do more assays with more timepoints, different concentrations. Maybe takes a certain [] to penetrate cell wall?

Also, restreaked kan-casette plates Made 3 ml cultures of each section (1 colony)


July 21, 2009

Restreakig of #2-4 After nearly four days of growth, I saw nothing but dead cells on the plates # 1 and # 5. So, I took what few white spots I saw on the plates # 2-4 and restreaked them.


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