User:Mariana Ruiz Velasco L./Notebook/IGEM 2010/Wet lab journal/2010/10/05

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GOSH!!!! Please help us!!!!

  • Today, after inactivating the enzyme, I ran a gel to check if everything went out well...

Rest mluc5Oct10.JPG

  • The lanes were as follows:

1. Ladder

2. 2-O in J23100(1)

3. 2-O in J23100(2)

4. 2-O in J23106(6)

5. 2-P in J23100(3)

6. 2-P in J23100(1)

7. 2-O in J23100(6)

  • Rethinking in my methodologies and experiments, I decided to ligate again the mutated luciferase with the J23102, so I ran a gel to quantify the parts for the ligation. I charged 3μl per sample, and 1.5μl for the plasmid. I also charged LRE both, with the DNA submission plasmid and the J23100 one.


  • So, the lanes are as follows:

1. Ladder

2. Mutated luciferase 2-O restricted with XBA I and PST I

3. Mutated luciferase 2-P restricted with XBA I and PST I

4, 5. LRE + pSB1C3 restricted with ECO RI and PST I

6. LRE + J23100 restricted with XBA I and PST I

7. Plasmid Bba_J61002 with dephosphated J23102.

  • By the end of the day, Melvin made the ligation of the restricted luciferase to both pSB1C3 and J23102, in hopes of having it ready to prove it on Tuesday. The reaction was as described previously.