User:Mariana Ruiz Velasco L./Notebook/IGEM 2010/Wet lab journal/2010/09/08

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It's been a hard day's night, and I'll be working like a dog
  - H2O -> 10  - Buffer 3 --> 2μL   - BSA > 1μL   - DNA > 5μL   - ECO RI > 1μL   - PST I --> 1μL  <br/ > <br/ > <br/ > <br/ > 1. Ladder.<br/ >
 * I started the day by extracting plasmid from the LRE + pSB1C3 strains so that I could check if the transformation was done correctly. After making a restriction for 20μL as follows, I ran a gel for 50min at 90V.<br/ >
 * The lanes were as follows and it can be observed that LRE from strains 3 and 5 was transformed successfully.<br/ >

2,3,4. LRE + pSB1C3.<br/ >

5. Green click beetle luciferase from purified PCR product.<br/ >

6. Red click beetle luciferase from purified PCR product.<br/ >

7. Mutated Photinus pyralis luciferase from purified PCR product.<br/ > <br/ > <br/ > <br/ > - H2O -> 13<br/ > <br/ > - Buffer 3 --> 3μL <br/ > <br/ > - BSA > 1μL <br/ > <br/ > - DNA > 10μL <br/ > <br/ > - XBA I -> 1.5μL <br/ > <br/ > - PST I --> 1.5μL<br/ > <br/ > <br/ > <br/ > <br/ >
 * About the click beetle experiment, I put a restriction for a total of 30μL so that I can be able to ligate it into a plasmid containing a constitutive promoter. The reaction was as follows and it was left incubating at 37°C ON:
 * Finally, for the luciferase + plasmid with T7 promoter, I inoculated 3 strains of it and 1 of the wildtype luciferase. Two out of the 3 pT7 strains were transformed and left incubating ON at 37°C.<br/ >


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