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

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July 7th Goal: Do restriction w/ Eco and BamHI

In +DNA... (buffer 3) NEB 10x = 1 ul 10x BSA = 1 ul ddH2O = 5 ul Eco/BamHI = 0.5 + 0.5 ul DNA = 2 ul Total = 10 ul

In -DNA... (buffer 3) NEB 10x = 1 ul 10x BSA = 1 ul ddH2O = 7 ul Eco/BamHI = 0.5 + 0.5 ul DNA = 0 ul Total = 10 ul

In -ENZ... (buffer 3) NEB 10x = 1 ul 10x BSA = 1 ul ddH2O = 6 ul Eco/BamHI = 0 ul DNA = 2 ul Total = 10 ul 37 degrees C, 1 hr, run on 1.2% agarose gel

Ran a gel

-Image insert here

1. Paraoxon came in. Review safety before opening

2. Assays will be 0.1 - 0.5 mM each

3. MW = 275.22 We have 100 mg or 0.000363 moles (0.363 mMoles)

100 mM solution...from 0.363 mMoles/3.63 ml = 0.1 mol / L solution

4. dilute in MeOH

5. store in Falcon tube or original sigma jar.

- use secondary containment (refer to M.Mattozzi)

6. helped Annie/Damon with their cultures of Aga2 display a)miniprepped Pcicen2 b) ran restriction digest

Check plates tomorrow


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