Week 14

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Revision as of 09:33, 8 October 2007 by Silvia.tamarri2 (Talk | contribs)
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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