Week 9

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Revision as of 10:53, 31 August 2007 by Francesca.bugane2 (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 1.2 ul Ligasi).
  • 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
  • We hold a meeting and the topics of the day are:

-model agreed;

-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 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.
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 I763012 + [http://partsregistry.org/Part:BBa_J06550 J06550] (I763016), I763012 + S0103 (missed in the sendbox),[http://partsregistry.org/Part:BBa_B0034 B0034] + J22101 (missed in the sendbox), [http://partsregistry.org/Part:BBa_B0034 B0034] + J04631 (I763020).
  • We transform the ligations.



08/30/07
  • There are colonies in all ligations.
  • We inoculate a 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.


08/31/07
  • Miniprep of I763012 + J06550 (I763016), I763012 + S0103 (missed in the sendbox), B0034 +

J22101 (missed in the sendbox), B0034 + J04631 (I763020), E0020 and of R0040.


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