User:Jorge E. Buendia Buendia/Notebook/iGEM UNAM-Genomics-Mexico/2010/06/21

iGEM UNAM-Genomics-Mexico

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June 21st, 2010 1. Transformation with backbone plasmid pSB3K3-J04450, 2009 Kit plate 1, well 7E. The DNA extracted from the plate was stored in a tube marked “pSB3K3-J04450 21.JUN.10”. I also made transformation with p17. 2. PCR to add minimum blue promoter to p30, I did the PCR reaction with rTth and Taq Platinum enzymes. I used plasmid 30 from the kit plate 1, distribution 2009, well 3M, I suspended the DNA from the plate in 10 ul of HPLC and used 2 ul of this suspension for each PCR reaction. As control I used 2 ul of Paz’s biopart D (~3 kb’s) diluted 1:5. I did two reactions for the plasmid 30 with the minimum blue promoter fro each enzyme and just one reaction for the control of 3 kb’s for each enzyme. 3. Run gel to verify PCR results. Lanes: 1,8) Ladder; 2) p30-minBP 1 rTth; 3) p30-minBP 2 rTth; 4) Ctrl 3 kb’s rTth; 5) p30-minBP 1 Platinum; 6) p30-minBP 2 Platinum; 7) Ctrl 3 kb’s Platinum. 4. Prepare culture dishes with LB medium and Tetracicline (Tet). Protocol to prepare them is explained on april 20th.