From 2007.igem.org

10/01/07

• Meeting: Ethical issue.

10/02/07

• Digestion of PSB4A3;
• Run on electrophoresis gel.
• pSB4A3 and Ptet-RFP band extraction.
• Ligation for

-pLac-GFP+I76303;

-I763012+I763020(I763032).

10/03/07

• Digestion of:

-pLac-Lacy-GFP with Eco/Pst1;

-I763031 with Eco/Pst1;

-I763036 with Eco/Pst1;

-pSB4A3 with Eco/Pst1.

• Run on electrophoresis gel.
• I763031, I763036 and pSB4A3 band extraction.
• Transformation of

-pTet-RFP-Plac GFP;

-I763031;

-I763032.

• Ligation for

-I763031+pSB4A3;

-I763032+pSB4A3.

Experimental Setup

• To have an higher precision for our measurements, we have decided to get, for the same field of view on the same sample of bacteria solution, either a short film segment (from which we can extract frames) and a Microsoft Excel file, which contains a ten seconds interval of values from the photomultiplier output, taken at a 100Hz frequency.

• In order to have the same focus for the microscope's oculars, the photocamera and the photomultiplier we have to calibrate our system by changing the position of both camera and photomultiplier.

• Finally, we check that the photomultiplier field of view is the same size as the microscope's one: by moving the appropiate diaphragms, we adjust the previously done settings.

10/04/07

• Trasformation of

-I763031+pSB4A3;

-I763032+pSB4A3.

• We inoculate I763034,I763031+pSB4A3 and I763032+pSB4A3 (in 6 ml).

10/05/07

• Stock preparation (500 ul) of I763034,I763031+pSB4A3 and I763032+pSB4A3.
• Miniprep of I763034,I763031+pSB4A3 and I763032+pSB4A3.
• Control digestion of

-I763034 with Xba/Pst1;

-I763031 with Eco/Xba and Xba/Pst1;

-I763032 with Eco/Xba and Xba/Pst1.

• Run on electrophoresis gel and I763032(Eco/Xba and Xba/Pst1) band extraction.

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