Week 5

From 2007.igem.org

(Difference between revisions)
Line 1: Line 1:
::'''07/30/07'''
::'''07/30/07'''
-
*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.
Line 28: Line 28:
-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;
Line 59: Line 59:
-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.
-
 
+
Line 72: Line 71:
- 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
  • 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].
  • 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).



07/31/07
  • 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 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.

-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 [http://partsregistry.org/Part:BBa_I52034 I52034] and [http://partsregistry.org/Part:BBa_B0015 B0015];

-Digestion of [http://partsregistry.org/Part:BBa_B0015 B0015] with Xba/Pst1;

-Digestion of [http://partsregistry.org/Part:BBa_I52034 I52034] with Spe/Pst1;

- 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_J04500 J04500]+[http://partsregistry.org/Part:BBa_C0051 C0051][http://partsregistry.org/Part:BBa_I763005 (I763005)]ligation which gives colonies so we inoculate in 5ml;

-Transformation for [http://partsregistry.org/Part:BBa_C0012 C0012] (4ul) and for [http://partsregistry.org/Part:BBa_J22101 J22101] (2ul);

-A new [http://partsregistry.org/Part:BBa_R0051 R0051]+ [http://partsregistry.org/Part:BBa_I13507 I13507] [http://partsregistry.org/Part:BBa_I763007 (I763007)] (vector 4ul, insert 8ul) ligation.



08/02/07
  • Today we do:

-Miniprep of [http://partsregistry.org/Part:BBa_J04500 J04500]+[http://partsregistry.org/Part:BBa_C0051 C0051] [http://partsregistry.org/Part:BBa_I763005 (I763005)];

-Control digestion of 5 ul with Eco/Pst1;

-A new [http://partsregistry.org/Part:BBa_B0015 B0015] digestion with Xba/Pst1;

-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];

-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;

-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 check if the RFP filter works properly.


08/03/07
  • We get along with:

-Miniprep for [http://partsregistry.org/Part:BBa_J22101 J22101] and for [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_J22101 J22101] digestion with Xba/Pst1 ;

- 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 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 stocks preparation;



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