Week 9

From 2007.igem.org

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Revision as of 14:04, 3 September 2007

08/27/07

New R0051+P0412 (I763010) ligation with a new protocol (3h 15°C and 1.2 ul Ligase).
Transformation of R0051+P0412 (I763010) ligation and of new biobricks S03520, S0100, B0034, J06550, S0103.

08/28/07

Lab meeting on:

-model -LacI cloning under the pTetR R0040)promoter control;

We transform new biobricks:E0020 and R0040

We inoculate a S03520, S0100, B0034, J06550, S0103 colony in 5 ml;

We observe that there are no colonies to R0051+P0412((I763010) ligation.

08/29/07

Miniprep of S03520, S0100, B0034, J06550, S0103.
S03520, S0100, J06550, S0103 digestions with Xba/Pst1.
B0034 digestion with Spe/Pst1.
Run on electroforesi gel and S03520, 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.
Ligation with a new protocol of I763012 + J06550 (I763016), I763012 + S0103 (missed in the sendbox),B0034 + J22101 (missed in the sendbox), B0034 + J04631 (I763020).
We transform the ligations.

08/30/07

There are colonies in the ligations.
We inoculate ligations, E0020 and 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 I763012 + J06550 (I763016), I763012+ S0103 (missed in the sandbox), B0034 +

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

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 ::'''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].
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 ::'''08/28/07'''
-*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 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'''
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 ::'''08/30/07'''
-*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'''
-*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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Week 9

From 2007.igem.org

Revision as of 14:04, 3 September 2007