Week 14

From 2007.igem.org

(Difference between revisions)
Line 5: Line 5:
::'''10/02/07'''
::'''10/02/07'''
-
*Digestion of PSB4A3;
+
*Digestion of [http://partsregistry.org/Part:pSB4A3 pSB4A3];
*Run on electrophoresis gel.
*Run on electrophoresis gel.
-
*pSB4A3 and I763033 band extraction.
+
*[http://partsregistry.org/Part:pSB4A3 pSB4A3]; and [http://partsregistry.org/Part:I763033 I763033] band extraction.
-
*Ligation for  
+
*Ligation for:
-
::-pLac-GFP+I76303;
+
::-[http://partsregistry.org/Part:I763004 I763004]+[http://partsregistry.org/Part:I763033 I763033];
-
::-I763012+I763020(I763032).
+
::-[http://partsregistry.org/Part:I763012 I763012]+[http://partsregistry.org/Part:I763020 I763020]([http://partsregistry.org/Part:I763032 (I763032)].
Line 17: Line 17:
::'''10/03/07'''
::'''10/03/07'''
*Digestion of:
*Digestion of:
-
::-I763032 with Eco/Pst1;
+
::-[http://partsregistry.org/Part:I763032 I763032] with Eco/Pst1;
-
::-I763031 with Eco/Pst1;
+
::-[http://partsregistry.org/Part:I763031 I763031] with Eco/Pst1;
-
::-I763036 with Eco/Pst1;
+
::-[http://partsregistry.org/Part:I763036 I763036] with Eco/Pst1;
-
::-pSB4A3 with Eco/Pst1.
+
::-[http://partsregistry.org/Part:pSB4A3 pSB4A3] with Eco/Pst1.
*Run on electrophoresis gel.
*Run on electrophoresis gel.
-
*I763031, I763036 and pSB4A3 band extraction.
+
*[http://partsregistry.org/Part:I763031 I763031], ]+[http://partsregistry.org/Part:I763036 I763036] and [http://partsregistry.org/Part:pSB4A3 pSB4A3] band extraction.
*Transformation of  
*Transformation of  
-
::-I763034;  
+
::-[http://partsregistry.org/Part:I763034 I763034];  
-
::-I763031;
+
::-[http://partsregistry.org/Part:I763031 I763031];
-
::-I763032.
+
::-[http://partsregistry.org/Part:I763032 I763032].
*Ligation for
*Ligation for
-
::-I763031+pSB4A3;
+
::-[http://partsregistry.org/Part:I763031 I763031]+[http://partsregistry.org/Part:pSB4A3 pSB4A3];
-
::-I763032+pSB4A3.
+
::-[http://partsregistry.org/Part:I763032 I763032::-[http://partsregistry.org/Part:I763032 I763032].
'''Experimental Setup'''
'''Experimental Setup'''
Line 43: Line 43:
::'''10/04/07'''
::'''10/04/07'''
*Trasformation of
*Trasformation of
-
::-I763031+pSB4A3;
+
::-[http://partsregistry.org/Part:I763031 I763031]+[http://partsregistry.org/Part:pSB4A3 pSB4A3];
-
::-I763032+pSB4A3.
+
::-[http://partsregistry.org/Part:I763032 I763032]+[http://partsregistry.org/Part:pSB4A3 pSB4A3];
-
*We inoculate I763034,I763031+pSB4A3 and I763032+pSB4A3 (in 6 ml).
+
*We inoculate [http://partsregistry.org/Part:I763034 I763034],I763031+pSB4A3 and [http://partsregistry.org/Part:I763032 I763032].+pSB4A3 (in 6 ml).

Revision as of 10:34, 8 October 2007

10/01/07
  • Meeting: Ethical issue.


10/02/07
  • Digestion of pSB4A3;
  • Run on electrophoresis gel.
  • pSB4A3; and I763033 band extraction.
  • Ligation for:
-I763004+I763033;
-I763012+I763020((I763032).



10/03/07
  • Digestion of:
-I763032 with Eco/Pst1;
-I763031 with Eco/Pst1;
-I763036 with Eco/Pst1;
-pSB4A3 with Eco/Pst1.
-I763034;
-I763031;
-I763032.
  • Ligation for
-I763031+pSB4A3;
-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
-I763031+pSB4A3;
-I763032+pSB4A3;



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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