Team:BYU Provo/Team OxyR/Week15

From 2011.igem.org

Team BYU Provo

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Indiana Trip
Outreach

25 July 2011

Matt set up overnight cultures of the strains (IGM13-21) to be used in the plate reader experiment on July 26th. Sat down with Mackay later and they decided that they didn't know which concentrations of Hydrogen Peroxide they would want to use to activate the OxyR promoters. After talking more, they realized they hadn't discussed a protocol yet with Dr. Grose, so they jotted down questions and went down to ask her. The resulting protocol was the precursor to the "Plate Reader Experiment" Protocol as now described. Some important points nailed down on July 25th: 1. In analyzing our data, we need to normalize with respect to OD. 2.The lethal concentration of H2O2 is above 1 mM. 3. The maximum oxyR-regulated response to H2O2 in a similar biosensor system is around 1 mM.

28 July 2011

Rob - ran colony PCR gel from 23 July.

Gel Printout

Matt determined a to-do list for the next few days:

Make more LB
Re-amplify OxyR-Medium fragment
Electroporate the plasmids with OxyR Weak, Medium, and Strong fragments into SoxR- and OxyR- strains.
Do Plate-reader experiment (using higher doses of H2O2 and measuring OD's)