Week 9

From 2007.igem.org

(Difference between revisions)
Line 1: Line 1:
::'''08/27/07'''
::'''08/27/07'''
-
*New [http://partsregistry.org/Part:BBa_R0051 R0051]+[http://partsregistry.org/Part:BBa_P0412 P0412] ([http://partsregistry.org/Part:BBa_I763010 I763010]) ligation with a new protocol (3h 15°C and 1.2 ul Ligasi).
+
*New [http://partsregistry.org/Part:BBa_R0051 R0051]+[http://partsregistry.org/Part:BBa_P0412 P0412] ([http://partsregistry.org/Part:BBa_I763010 I763010]) ligation with a new protocol (3h 15°C and 1.2 ul Ligase).
*Transformation of [http://partsregistry.org/Part:BBa_R0051 R0051]+[http://partsregistry.org/Part:BBa_P0412 P0412] ([http://partsregistry.org/Part:BBa_I763010 I763010]) ligation and of new biobricks [http://partsregistry.org/Part:BBa_S03520 S03520], [http://partsregistry.org/Part:BBa_S0100 S0100], [http://partsregistry.org/Part:BBa_B0034 B0034], [http://partsregistry.org/Part:BBa_J06550 J06550], [http://partsregistry.org/Part:BBa_S0103 S0103].  
*Transformation of [http://partsregistry.org/Part:BBa_R0051 R0051]+[http://partsregistry.org/Part:BBa_P0412 P0412] ([http://partsregistry.org/Part:BBa_I763010 I763010]) ligation and of new biobricks [http://partsregistry.org/Part:BBa_S03520 S03520], [http://partsregistry.org/Part:BBa_S0100 S0100], [http://partsregistry.org/Part:BBa_B0034 B0034], [http://partsregistry.org/Part:BBa_J06550 J06550], [http://partsregistry.org/Part:BBa_S0103 S0103].  
Line 7: Line 7:
::'''08/28/07'''
::'''08/28/07'''
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*We hold a meeting and the topics of the day are:
+
*Lab meeting on: 
 +
-model
 +
-LacI cloning under the pTetR [http://partsregistry.org/Part:BBa_R0040 R0040])promoter control;
-
-model agreed;
+
*We transform new biobricks:[http://partsregistry.org/Part:BBa_E0020 E0020] and [http://partsregistry.org/Part:BBa_R0040 R0040]
-
 
+
-
-we agree the position of LacI not controlled by Plac but by costitutionaly expressed as PTetR gene ([http://partsregistry.org/Part:BBa_R0040 R0040]);
+
-
 
+
-
-reseach of genomic pLacI sequence to draw the primers or to order the synthesis.
+
-
 
+
-
*We transform new biobricks:[http://partsregistry.org/Part:BBa_E0020 E0020] (to put after CMV [http://partsregistry.org/Part:BBa_J52034 J52034] promoter) and [http://partsregistry.org/Part:BBa_R0040 R0040] (whith witch adjust LacI expression);
+
* We inoculate a [http://partsregistry.org/Part:BBa_S03520 S03520], [http://partsregistry.org/Part:BBa_S0100 S0100], [http://partsregistry.org/Part:BBa_B0034 B0034], [http://partsregistry.org/Part:BBa_J06550 J06550], [http://partsregistry.org/Part:BBa_S0103 S0103] colony in 5 ml;
* We inoculate a [http://partsregistry.org/Part:BBa_S03520 S03520], [http://partsregistry.org/Part:BBa_S0100 S0100], [http://partsregistry.org/Part:BBa_B0034 B0034], [http://partsregistry.org/Part:BBa_J06550 J06550], [http://partsregistry.org/Part:BBa_S0103 S0103] colony in 5 ml;
-
* We observe that there isn't a colony to [http://partsregistry.org/Part:BBa_R0051 R0051]+[http://partsregistry.org/Part:BBa_P0412 P0412]([http://partsregistry.org/Part:BBa_I763010 (I763010)] ligation, so we abandon this way.
+
* We observe that there are no colonies to [http://partsregistry.org/Part:BBa_R0051 R0051]+[http://partsregistry.org/Part:BBa_P0412 P0412]([http://partsregistry.org/Part:BBa_I763010 (I763010)] ligation.
::'''08/29/07'''
::'''08/29/07'''
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::'''08/30/07'''
::'''08/30/07'''
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*There are colonies in all ligations.
+
*There are colonies in the ligations.
-
*We inoculate a ligations, [http://partsregistry.org/Part:BBa_E0020 E0020] and [http://partsregistry.org/Part:BBa_R0040 R0040] colony in 5 ml.
+
*We inoculate ligations, [http://partsregistry.org/Part:BBa_E0020 E0020] and [http://partsregistry.org/Part:BBa_R0040 R0040] colony in 5 ml.
-
*Dimension tests with PMT to set gain and dimension area.
+
*We perform some tests to set up the fluorescence acquisition with the PMT (photomultiplier tube).
      
      
::'''08/31/07'''
::'''08/31/07'''
-
*Miniprep of [http://partsregistry.org/Part:BBa_I763012 I763012] + [http://partsregistry.org/Part:BBa_J06550 J06550] [http://partsregistry.org/Part:BBa_I763016 (I763016)], [http://partsregistry.org/Part:BBa_I763012 I763012]+ [http://partsregistry.org/Part:BBa_S0103 S0103] (missed in the sendbox), [http://partsregistry.org/Part:BBa_B0034 B0034] +   
+
*Miniprep of [http://partsregistry.org/Part:BBa_I763012 I763012] + [http://partsregistry.org/Part:BBa_J06550 J06550] [http://partsregistry.org/Part:BBa_I763016 (I763016)], [http://partsregistry.org/Part:BBa_I763012 I763012]+ [http://partsregistry.org/Part:BBa_S0103 S0103] (missed in the sandbox), [http://partsregistry.org/Part:BBa_B0034 B0034] +   
-
[http://partsregistry.org/Part:BBa_J22101 J22101] (missed in the sendbox), [http://partsregistry.org/Part:BBa_B0034 B0034] + [http://partsregistry.org/Part:BBa_J04631 J04631] [http://partsregistry.org/Part:BBa_I763020 (I763020)], [http://partsregistry.org/Part:BBa_E0020  E0020 ] and of [http://partsregistry.org/Part:BBa_R0040 R0040].
+
[http://partsregistry.org/Part:BBa_J22101 J22101] (missed in the sandbox), [http://partsregistry.org/Part:BBa_B0034 B0034] + [http://partsregistry.org/Part:BBa_J04631 J04631] [http://partsregistry.org/Part:BBa_I763020 (I763020)], [http://partsregistry.org/Part:BBa_E0020  E0020 ] and of [http://partsregistry.org/Part:BBa_R0040 R0040].
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Revision as of 14:04, 3 September 2007

08/27/07
  • New [http://partsregistry.org/Part:BBa_R0051 R0051]+[http://partsregistry.org/Part:BBa_P0412 P0412] ([http://partsregistry.org/Part:BBa_I763010 I763010]) ligation with a new protocol (3h 15°C and 1.2 ul Ligase).
  • Transformation of [http://partsregistry.org/Part:BBa_R0051 R0051]+[http://partsregistry.org/Part:BBa_P0412 P0412] ([http://partsregistry.org/Part:BBa_I763010 I763010]) ligation and of new biobricks [http://partsregistry.org/Part:BBa_S03520 S03520], [http://partsregistry.org/Part:BBa_S0100 S0100], [http://partsregistry.org/Part:BBa_B0034 B0034], [http://partsregistry.org/Part:BBa_J06550 J06550], [http://partsregistry.org/Part:BBa_S0103 S0103].



08/28/07
  • Lab meeting on:

-model -LacI cloning under the pTetR [http://partsregistry.org/Part:BBa_R0040 R0040])promoter control;

  • We transform new biobricks:[http://partsregistry.org/Part:BBa_E0020 E0020] and [http://partsregistry.org/Part:BBa_R0040 R0040]
  • We inoculate a [http://partsregistry.org/Part:BBa_S03520 S03520], [http://partsregistry.org/Part:BBa_S0100 S0100], [http://partsregistry.org/Part:BBa_B0034 B0034], [http://partsregistry.org/Part:BBa_J06550 J06550], [http://partsregistry.org/Part:BBa_S0103 S0103] colony in 5 ml;
  • We observe that there are no colonies to [http://partsregistry.org/Part:BBa_R0051 R0051]+[http://partsregistry.org/Part:BBa_P0412 P0412]([http://partsregistry.org/Part:BBa_I763010 (I763010)] ligation.
08/29/07
  • Miniprep of [http://partsregistry.org/Part:BBa_S03520 S03520], [http://partsregistry.org/Part:BBa_S0100 S0100], [http://partsregistry.org/Part:BBa_B0034 B0034], [http://partsregistry.org/Part:BBa_J06550 J06550], [http://partsregistry.org/Part:BBa_S0103 S0103].
  • [http://partsregistry.org/Part:BBa_S03520 S03520], [http://partsregistry.org/Part:BBa_S0100 S0100], [http://partsregistry.org/Part:BBa_J06550 J06550], [http://partsregistry.org/Part:BBa_S0103 S0103] digestions with Xba/Pst1.
  • [http://partsregistry.org/Part:BBa_B0034 B0034] digestion with Spe/Pst1.
  • Run on electroforesi gel and [http://partsregistry.org/Part:BBa_S03520 S03520], [http://partsregistry.org/Part:BBa_S0100 S0100], [http://partsregistry.org/Part:BBa_B0034 B0034] band extraction. We notice problems with [http://partsregistry.org/Part:BBa_J06550 J06550] band height (the insert is too heavy) and with [http://partsregistry.org/Part:BBa_S0103 S0103] band because the plasmid too heavy, however we extract insert bands for both.
  • Ligation with a new protocol of [http://partsregistry.org/Part:BBa_I763012 I763012] + [http://partsregistry.org/Part:BBa_J06550 J06550] ([http://partsregistry.org/Part:BBa_I763016 I763016]), [http://partsregistry.org/Part:BBa_I763012 I763012] + [http://partsregistry.org/Part:BBa_S0103 S0103] (missed in the sendbox),[http://partsregistry.org/Part:BBa_B0034 B0034] + [http://partsregistry.org/Part:BBa_J22101 J22101] (missed in the sendbox), [http://partsregistry.org/Part:BBa_B0034 B0034] + [http://partsregistry.org/Part:BBa_J04631 J04631] ([http://partsregistry.org/Part:BBa_I763020 I763020]).
  • We transform the ligations.



08/30/07
  • There are colonies in the ligations.
  • We inoculate ligations, [http://partsregistry.org/Part:BBa_E0020 E0020] and [http://partsregistry.org/Part:BBa_R0040 R0040] colony in 5 ml.
  • We perform some tests to set up the fluorescence acquisition with the PMT (photomultiplier tube).


08/31/07
  • Miniprep of [http://partsregistry.org/Part:BBa_I763012 I763012] + [http://partsregistry.org/Part:BBa_J06550 J06550] [http://partsregistry.org/Part:BBa_I763016 (I763016)], [http://partsregistry.org/Part:BBa_I763012 I763012]+ [http://partsregistry.org/Part:BBa_S0103 S0103] (missed in the sandbox), [http://partsregistry.org/Part:BBa_B0034 B0034] +

[http://partsregistry.org/Part:BBa_J22101 J22101] (missed in the sandbox), [http://partsregistry.org/Part:BBa_B0034 B0034] + [http://partsregistry.org/Part:BBa_J04631 J04631] [http://partsregistry.org/Part:BBa_I763020 (I763020)], [http://partsregistry.org/Part:BBa_E0020 E0020 ] and of [http://partsregistry.org/Part:BBa_R0040 R0040].


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