Week 5

From 2007.igem.org

(Difference between revisions)
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::'''07/30/07'''
::'''07/30/07'''
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*We control bacterial growth curve (link).
+
*We check bacterial growth curve (link).
*Transformation for [http://partsregistry.org/Part:BBa_C0012 C0012] and [http://partsregistry.org/Part:BBa_J52034 J52034] and [http://partsregistry.org/Part:BBa_B0015 B0015].
*Transformation for [http://partsregistry.org/Part:BBa_C0012 C0012] and [http://partsregistry.org/Part:BBa_J52034 J52034] and [http://partsregistry.org/Part:BBa_B0015 B0015].
*We get along with the ligation [http://partsregistry.org/Part:BBa_R0051 R0051]+[http://partsregistry.org/Part:BBa_I13507 I13507] [http://partsregistry.org/Part:BBa_I763007 (I763007)] (vector 2 ul, insert 6 ul) and the ligation [http://partsregistry.org/Part:BBa_J04500 J04500]+ [http://partsregistry.org/Part:BBa_C0051 C0051] [http://partsregistry.org/Part:BBa_I763005 (I763005)](vector 2 ul, insert 10 ul).
*We get along with the ligation [http://partsregistry.org/Part:BBa_R0051 R0051]+[http://partsregistry.org/Part:BBa_I13507 I13507] [http://partsregistry.org/Part:BBa_I763007 (I763007)] (vector 2 ul, insert 6 ul) and the ligation [http://partsregistry.org/Part:BBa_J04500 J04500]+ [http://partsregistry.org/Part:BBa_C0051 C0051] [http://partsregistry.org/Part:BBa_I763005 (I763005)](vector 2 ul, insert 10 ul).
Line 8: Line 8:
::'''07/31/07'''
::'''07/31/07'''
-
*There isn’t any colony for [http://partsregistry.org/Part:BBa_C0012 C0012] , so we inoculate [http://partsregistry.org/Part:BBa_I52034 I52034] and [http://partsregistry.org/Part:BBa_B0015 B0015] colony in 5 ml.
+
*There are no colonies for [http://partsregistry.org/Part:BBa_C0012 C0012] , so we inoculate [http://partsregistry.org/Part:BBa_I52034 I52034] and [http://partsregistry.org/Part:BBa_B0015 B0015] colony in 5 ml.
*We strake on plates 30/07 ligations.
*We strake on plates 30/07 ligations.
*We get along with the [http://partsregistry.org/Part:BBa_J04431 J04431] fluorescence test:
*We get along with the [http://partsregistry.org/Part:BBa_J04431 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.
+
-Every 15 minutes we check bacterial OD and fluorescence. Cells are fluorescent at t= 210 min and 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 decide to repeat the test to understand if we can switch off the fluorescent cells with a higher amount of glucose.
-
We will attend this test on 07/08.
+
We will perform this test on 07/08.
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-Digestion of [http://partsregistry.org/Part:BBa_I52034 I52034] with Spe/Pst1;
-Digestion of [http://partsregistry.org/Part:BBa_I52034 I52034] with Spe/Pst1;
-
-Extraction from gel (we can’t see [http://partsregistry.org/Part:BBa_B0015 B0015]);
+
- Gel extraction (we can’t see [http://partsregistry.org/Part:BBa_B0015 B0015]);
-[http://partsregistry.org/Part:BBa_R0015 R0015]+[http://partsregistry.org/Part:BBa_I13507 I13507][http://partsregistry.org/Part:BBa_I763007 (I763007)] ligation which doesn’t give colonies so we transform with the other 10 ul of ligation;  
-[http://partsregistry.org/Part:BBa_R0015 R0015]+[http://partsregistry.org/Part:BBa_I13507 I13507][http://partsregistry.org/Part:BBa_I763007 (I763007)] ligation which doesn’t give colonies so we transform with the other 10 ul of ligation;  
Line 49: Line 49:
-A new [http://partsregistry.org/Part:BBa_B0015 B0015] digestion with Xba/Pst1;
-A new [http://partsregistry.org/Part:BBa_B0015 B0015] digestion with Xba/Pst1;
-
-Digested run on electroforesi gel but we have problems with gel because we don’t see the band;
+
-Run on electroforesis gel. We have problems with gel because we don’t see the band;
-Band extraction for [http://partsregistry.org/Part:BBa_B0015 B0015] e [http://partsregistry.org/Part:BBa_I763005 I763005];
-Band extraction for [http://partsregistry.org/Part:BBa_B0015 B0015] e [http://partsregistry.org/Part:BBa_I763005 I763005];
-
-We observe there aren’t colonies for [http://partsregistry.org/Part:BBa_C0012 C0012], but there are colonies for [http://partsregistry.org/Part:BBa_R0051 R0051] + [http://partsregistry.org/Part:BBa_I13507 I13507][http://partsregistry.org/Part:BBa_I763007 (I763007)], so we identify the correct ligation protocol;
+
-There are no colonies for [http://partsregistry.org/Part:BBa_C0012 C0012], but there are colonies for [http://partsregistry.org/Part:BBa_R0051 R0051] + [http://partsregistry.org/Part:BBa_I13507 I13507][http://partsregistry.org/Part:BBa_I763007 (I763007)]. So, we identify the correct ligation protocol;
- [http://partsregistry.org/Part:BBa_R0010 R0010], [http://partsregistry.org/Part:BBa_P0412 P0412], [http://partsregistry.org/Part:BBa_C0012 C0012]  transformation;
- [http://partsregistry.org/Part:BBa_R0010 R0010], [http://partsregistry.org/Part:BBa_P0412 P0412], [http://partsregistry.org/Part:BBa_C0012 C0012]  transformation;
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-Inoculation in 5 ml for [http://partsregistry.org/Part:BBa_R0051 R0051] + [http://partsregistry.org/Part:BBa_I13507 I13507] and for [http://partsregistry.org/Part:BBa_J22101 J22101];
-Inoculation in 5 ml for [http://partsregistry.org/Part:BBa_R0051 R0051] + [http://partsregistry.org/Part:BBa_I13507 I13507] and for [http://partsregistry.org/Part:BBa_J22101 J22101];
-
- [http://partsregistry.org/Part:BBa_R0051 R0051] + [http://partsregistry.org/Part:BBa_I13507 I13507] [http://partsregistry.org/Part:BBa_I763007 (I763007)]  fluorescence test to verify if the filter for RFP is adapted, but it isn’t.
+
- [http://partsregistry.org/Part:BBa_R0051 R0051] + [http://partsregistry.org/Part:BBa_I13507 I13507] [http://partsregistry.org/Part:BBa_I763007 (I763007)]  fluorescence test to check if the RFP filter works properly.
-
 
+
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- Control digestion of [http://partsregistry.org/Part:BBa_R0051 R0051] + [http://partsregistry.org/Part:BBa_I13507 I13507]  [http://partsregistry.org/Part:BBa_I763007 (I763007)];
- Control digestion of [http://partsregistry.org/Part:BBa_R0051 R0051] + [http://partsregistry.org/Part:BBa_I13507 I13507]  [http://partsregistry.org/Part:BBa_I763007 (I763007)];
-
- [http://partsregistry.org/Part:BBa_J04500 J04500]+[http://partsregistry.org/Part:BBa_C0051 C0051] [http://partsregistry.org/Part:BBa_I763005 (I763005)] and [http://partsregistry.org/Part:BBa_B0015 B0015]and [http://partsregistry.org/Part:BBa_J22101 J22101] and [http://partsregistry.org/Part:BBa_R0051 R0051] + [http://partsregistry.org/Part:BBa_I13507 I13507] [http://partsregistry.org/Part:BBa_I763007 (I763007)] run on electroforesi gel and [http://partsregistry.org/Part:BBa_J04500 J04500]+[http://partsregistry.org/Part:BBa_C0051 C0051] [http://partsregistry.org/Part:BBa_I763005 (I763005)] and  [http://partsregistry.org/Part:BBa_J22101 J22101] estraction;
+
- [http://partsregistry.org/Part:BBa_J04500 J04500]+[http://partsregistry.org/Part:BBa_C0051 C0051] [http://partsregistry.org/Part:BBa_I763005 (I763005)] and [http://partsregistry.org/Part:BBa_B0015 B0015]and [http://partsregistry.org/Part:BBa_J22101 J22101] and [http://partsregistry.org/Part:BBa_R0051 R0051] + [http://partsregistry.org/Part:BBa_I13507 I13507] [http://partsregistry.org/Part:BBa_I763007 (I763007)] run on electroforesis gel and [http://partsregistry.org/Part:BBa_J04500 J04500]+[http://partsregistry.org/Part:BBa_C0051 C0051] [http://partsregistry.org/Part:BBa_I763005 (I763005)] and  [http://partsregistry.org/Part:BBa_J22101 J22101] extraction;
-
-Glycerol stock preparation;
+
-Glycerol stocks preparation;

Revision as of 13:45, 3 September 2007

07/30/07



07/31/07
  • There are no colonies for C0012 , so we inoculate I52034 and B0015 colony in 5 ml.
  • We strake on plates 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.

-Every 15 minutes we check bacterial OD and fluorescence. Cells are fluorescent at t= 210 min and OD=0.5.

  • We decide to repeat the test to understand if we can switch off the fluorescent cells with a higher amount of glucose.

We will perform this test on 07/08.



08/01/07
  • We get along with:

-Miniprep of I52034 and B0015;

-Digestion of B0015 with Xba/Pst1;

-Digestion of I52034 with Spe/Pst1;

- Gel extraction (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
  • Today we do:

-Miniprep of J04500+C0051 (I763005);

-Control digestion of 5 ul with Eco/Pst1;

-A new B0015 digestion with Xba/Pst1;

-Run on electroforesis gel. We have problems with gel because we don’t see the band;

-Band extraction for B0015 e I763005;

-There are no colonies for C0012, but there are colonies for R0051 + I13507(I763007). So, we identify the correct ligation protocol;

- R0010, P0412, C0012 transformation;

-Inoculation in 5 ml for R0051 + I13507 and for J22101;

- R0051 + I13507 (I763007) fluorescence test to check if the RFP filter works properly.


08/03/07
  • We get along with:

-Miniprep for J22101 and for R0051+ I13507 (I763007);

- J22101 digestion with Xba/Pst1 ;

- Control digestion of R0051 + I13507 (I763007);

- J04500+C0051 (I763005) and B0015and J22101 and R0051 + I13507 (I763007) run on electroforesis gel and J04500+C0051 (I763005) and J22101 extraction;

-Glycerol stocks preparation;



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