Week 5

From 2007.igem.org

(Difference between revisions)
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*We get along with the J04431 fluorescence test:
*We get along with the J04431 fluorescence test:
-From the glycerol stock we sprout 1 ml at 37°C for 1 hour in agitation.
-From the glycerol stock we sprout 1 ml at 37°C for 1 hour in agitation.
 +
-We verify the OD and the fluorescence every 15 minutes. The fluorescence compare at the istance t=210min and at the OD=0.5.
-We verify the OD and the fluorescence every 15 minutes. The fluorescence compare at the istance t=210min and at the OD=0.5.
*At the end of this test we decide to repeat it to see if we can switch off the fluorescent cells with a major quantity of glucose to confirm our assumptions.
*At the end of this test we decide to repeat it to see if we can switch off the fluorescent cells with a major quantity of glucose to confirm our assumptions.
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*We get along with:
*We get along with:
-Miniprep of I52034 and B0015;
-Miniprep of I52034 and B0015;
 +
-Digestion of B0015 with Xba/Pst1
-Digestion of B0015 with Xba/Pst1
                       I52034 with Spe/Pst1;
                       I52034 with Spe/Pst1;
 +
-extraction from gel (we can’t see B0015);
-extraction from gel (we can’t see B0015);
 +
-R0015+I13507 (I763007) ligation which doesn’t give colonies so we transform with the other 10 ul of ligation;  
-R0015+I13507 (I763007) ligation which doesn’t give colonies so we transform with the other 10 ul of ligation;  
 +
-J04500+C0051 (I763005) ligation which gives colonies so we inoculate in 5ml;
-J04500+C0051 (I763005) ligation which gives colonies so we inoculate in 5ml;
 +
-transformation for C0012 (4ul) and for J22101 (2ul);
-transformation for C0012 (4ul) and for J22101 (2ul);
 +
-a new R0051+ I13507 (I763007) (vector 4ul, insert 8ul) ligation.  
-a new R0051+ I13507 (I763007) (vector 4ul, insert 8ul) ligation.  

Revision as of 12:26, 27 August 2007

07/30/07
  • We control bacterial grew (link).
  • Transformation for C0012 and J52034 and B0015.
  • We get along with the ligation R0051+I13507 (I763007) (vector 2 ul, insert 6 ul) and the ligation J04500+ C0051 (I763005) (vector 2 ul, insert 10 ul).



07/31/07
  • There isn’t any colony for C0012, so we inoculate I52034 and B0015 colony in 5 ml.
  • We plate 30/07 ligations.
  • We get along with the J04431 fluorescence test:

-From the glycerol stock we sprout 1 ml at 37°C for 1 hour in agitation.

-We verify the OD and the fluorescence every 15 minutes. The fluorescence compare at the istance t=210min and at the OD=0.5.

  • At the end of this test we decide to repeat it to see if we can switch off the fluorescent cells with a major quantity of glucose to confirm our assumptions.

We will attend this test on 07/08.



08/01/07
  • We get along with:

-Miniprep of I52034 and B0015;

-Digestion of B0015 with Xba/Pst1 

                     I52034 with Spe/Pst1;

-extraction from gel (we can’t see B0015);

-R0015+I13507 (I763007) ligation which doesn’t give colonies so we transform with the other 10 ul of ligation;

-J04500+C0051 (I763005) ligation which gives colonies so we inoculate in 5ml;

-transformation for C0012 (4ul) and for J22101 (2ul);

-a new R0051+ I13507 (I763007) (vector 4ul, insert 8ul) ligation.



08/02/07



08/03/07



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