Week 9

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Revision as of 21:02, 7 September 2007

08/27/07

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

08/28/07

• Lab meeting on:

-model;

-LacI cloning under the pTetR R0040)promoter control.

• We transform new biobricks:E0020 and R0040

• We inoculate S03520, S0100, B0034, J06550, S0103 colonies in 5ml;

• There are no colonies for 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 electrophoresis gel and S03520, S0100, B0034 band extraction. We have some problems with J06550 plasmid vector due to its molecular weight (too heavy) and with S0103 insert for the same reason (too heavy). Thus, we decide to use another biobrick.
• Ligation for 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 07/30/07 plates.
• We inoculate a E0020 and R0040 colony in 5ml.
• We perform some tests to set up the fluorescence acquisition with the PMT (photomultiplier tube).

08/31/07

• Miniprep of :

-I763012 + J06550

-I763012+ S0103 (I763016);

-B0034 + J22101 (missed in the sandbox);

-B0034 + J04631 (I763020);

-E0022 ;

-R0040.

• Digestion for:

-I763012 + S0103 with Spe/Pst1;

-B0034 + J22101 with Xba/Pst1;

-B0034 + J04631 (I763020) with Xba/Pst1;

-E0022 with Xba/Pst1 ;

-R0040 with Spe/Pst1.

• Band extraction from gel for:

-I763012+ S0103 ((I763016);

-B0034 + J22101 (missed in the sandbox);

-B0034 + J04631 (I763020);

-E0020 ;

-R0040.

• Ligations for

-I763016+I763020;

-R0040+S0100;

-R0040+S03520;

-I763005+I763015;

-J52034+E0022;

• Transformations for the ligations.

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