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

From OpenWetWare
Jump to: navigation, search
Igem-logo-150px.png iGEM UNAM-Genomics-Mexico <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>      </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

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.

PCR p30-minBP 21Jun2010.JPG

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.