Week 9

From 2007.igem.org

(Difference between revisions)
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::'''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 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],
+
*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].
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::'''08/29/07'''
::'''08/29/07'''
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*Miniprep of [http://partsregistry.org/Part:BBa_S03520 S03520], [http://partsregistry.org/Part:BBa_S0100 S0100], B0034, J06550, S0103.
+
*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], J06550, S0103 digestions with Xba/Pst1.
+
*[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.
-
*B0034 digestion with Spe/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], B0034 band extraction. We notice problems with J06550 band height (the insert is too heavy) and with S0103 band because the plasmid too heavy, however we extract insert bands for both.
+
*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 + J06550 (I763016), I763012 + S0103 (missed in the sendbox), B0034 + J22101 (missed in the sendbox), B0034 + J04631 (I763020).
+
*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.
*We transform the ligations.

Revision as of 10:53, 31 August 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 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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