Week 9

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Revision as of 17:05, 9 September 2007 by Silvia.tamarri2 (Talk | contribs)
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 1.2ul Ligase).
  • We transform [http://partsregistry.org/Part:BBa_R0051 R0051]+[http://partsregistry.org/Part:BBa_P0412 P0412] ([http://partsregistry.org/Part:BBa_I763010 I763010]) ligation and new [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] biobricks.



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 [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] colonies in 5ml;
  • There are no colonies for [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 electrophoresis 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 have some problems with [http://partsregistry.org/Part:BBa_J06550 J06550] plasmid vector due to its molecular weight (too heavy) and with [http://partsregistry.org/Part:BBa_S0103 S0103] insert for the same reason (too heavy). Thus, we decide to use another biobrick.
  • Ligation for [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 07/30/07 plates.
  • We inoculate a [http://partsregistry.org/Part:BBa_E0020 E0020] and [http://partsregistry.org/Part:BBa_R0040 R0040] colony in 5ml.
  • 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_I763012 I763012]+ [http://partsregistry.org/Part:BBa_S0103 S0103] [http://partsregistry.org/Part:BBa_I763016 (I763016)];

-[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_E0022 E0022 ];

-[http://partsregistry.org/Part:BBa_R0040 R0040].

  • Digestion for:

-[http://partsregistry.org/Part:BBa_I763012 I763012] + [http://partsregistry.org/Part:BBa_S0103 S0103] with Spe/Pst1;

-[http://partsregistry.org/Part:BBa_B0034 B0034] + [http://partsregistry.org/Part:BBa_J22101 J22101] with Xba/Pst1;

-[http://partsregistry.org/Part:BBa_B0034 B0034] + [http://partsregistry.org/Part:BBa_J04631 J04631] [http://partsregistry.org/Part:BBa_I763020 (I763020)] with Xba/Pst1;

-[http://partsregistry.org/Part:BBa_E0022 E0022] with Xba/Pst1 ;

-[http://partsregistry.org/Part:BBa_R0040 R0040] with Spe/Pst1.

  • Band extraction from gel for:

-[http://partsregistry.org/Part:BBa_I763012 I763012]+ [http://partsregistry.org/Part:BBa_S0103 S0103] ([http://partsregistry.org/Part:BBa_I763016 (I763016)];

-[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 ];

-[http://partsregistry.org/Part:BBa_R0040 R0040].

  • Ligations at 15°C for 3h for:

-[http://partsregistry.org/Part:BBa_I763016 I763016]+[http://partsregistry.org/Part:BBa_I763020 I763020];

-[http://partsregistry.org/Part:BBa_R0040 R0040]+[http://partsregistry.org/Part:BBa_S0100 S0100];

-[http://partsregistry.org/Part:BBa_R0040 R0040]+[http://partsregistry.org/Part:BBa_S03520 S03520];

-[http://partsregistry.org/Part:BBa_I763005 I763005]+[http://partsregistry.org/Part:BBa_I763015 I763015];

-[http://partsregistry.org/Part:BBa_J52034 J52034]+[http://partsregistry.org/Part:BBa_E0022 E0022];

  • Transformation of all ligations.
  • We find colonies for all ligations, except for the first ligation.


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