User:Mariana Ruiz Velasco L./Notebook/IGEM 2010/Wet lab journal/2010/06/24

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 * style="background-color: #EEE"|[[Image:Logo_light_minimal_full_res.jpg|150px]] WiFi coli: A communicolight system
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Woohoo!!!!
     1. Ladder.  2. Luciferase with the mutation (complete).  3. Positive control.  
 * Today, I ran a gel to check if my PCR product was there:
 * The lanes were as follows:

Another PCR
  --> Mix 1 for 30 μL <--   -H2O > 9μl  -Buffer 3.3 -> 6μl <br/ > -Mg(OAc)2 -> 3μl <br/ > -dNTP's --> 4μl <br/ > -Prefix > 3μl <br/ > -Suffix > 3μl <br/ > -DNA > 2μl <br/ > <br/ > <br/ > --> Mix 2 for 20 μL <-- (this reaction will be added after the hot start) <br/ > <br/ > -H2O -> 10.5μl <br/ > -Buffer 3.3 --> 9μl <br/ > -rtTh polymerase --> 0.5μl <br/ > <br/ >
 * After the success, I went to see Miguel to check the next steps. This involves doing a PCR with a 5/100 DNA dilution (of the previous PCR product) using rtTh. The reaction is defined as follows and I did two reactions with the dilution and the respective controls.

<br/ > <br/ > <br/ > <br/ > 1. Ladder. <br/ > 2-7. Jorge's experiment. <br/ > 8. Negative control.<br/ > 9. Positive control.<br/ > 10 and 11. Mutated luciferase PCR product.<br/ > <br/ >
 * The gel (ran for 50min at 90V) shows that the two expected products are there:
 * The lanes are:


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