From 2007.igem.org

08/27/07

• New R0051+P0412 (I763010) ligation with a new protocol (3h 15°C and 1.2 ul Ligasi).
• Transformation of R0051+P0412 (I763010) ligation and of new biobricks S03520, S0100, B0034, J06550, 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 (R0040);

-reseach of genomic pLacI sequence to draw the primers or to order the synthesis.

• We transform new biobricks:E0020 (to put after CMV J52034 promoter) and R0040 (whith witch adjust LacI expression);

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

• We observe that there isn't a colony to R0051+P0412((I763010) ligation, so we abandon this way.

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 all ligations.
• We inoculate a ligations, E0020 and 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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