Week 14

From 2007.igem.org

Revision as of 10:36, 8 October 2007

10/01/07

• Meeting: Ethical issue.

10/02/07

• Digestion of [http://partsregistry.org/Part:pSB4A3 pSB4A3];
• Run on electrophoresis gel.
• [http://partsregistry.org/Part:pSB4A3 pSB4A3]; and [http://partsregistry.org/Part:I763033 I763033] band extraction.
• Ligation for:

-[http://partsregistry.org/Part:I763004 I763004]+[http://partsregistry.org/Part:I763033 I763033];

-[http://partsregistry.org/Part:I763012 I763012]+[http://partsregistry.org/Part:I763020 I763020][http://partsregistry.org/Part:I763032 (I763032)].

10/03/07

• Digestions of:

-[http://partsregistry.org/Part:I763032 I763032] with Eco/Pst1;

-[http://partsregistry.org/Part:I763031 I763031] with Eco/Pst1;

-[http://partsregistry.org/Part:I763036 I763036] with Eco/Pst1;

-[http://partsregistry.org/Part:pSB4A3 pSB4A3] with Eco/Pst1.

• Run on electrophoresis gel.
• [http://partsregistry.org/Part:I763031 I763031], [http://partsregistry.org/Part:I763036 I763036] and [http://partsregistry.org/Part:pSB4A3 pSB4A3] band extraction.
• Transformations of:

-[http://partsregistry.org/Part:I763034 I763034];

-[http://partsregistry.org/Part:I763031 I763031];

-[http://partsregistry.org/Part:I763032 I763032].

• Ligations for:

-[http://partsregistry.org/Part:I763031 I763031]+[http://partsregistry.org/Part:pSB4A3 pSB4A3];

-[http://partsregistry.org/Part:I763032 I763032::-[http://partsregistry.org/Part:I763032 I763032].

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

-[http://partsregistry.org/Part:I763031 I763031]+[http://partsregistry.org/Part:pSB4A3 pSB4A3];

-[http://partsregistry.org/Part:I763032 I763032]+[http://partsregistry.org/Part:pSB4A3 pSB4A3];

• We inoculate [http://partsregistry.org/Part:I763034 I763034],I763031+pSB4A3 and [http://partsregistry.org/Part:I763032 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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