User:Mariana Ruiz Velasco L./Notebook/IGEM 2010/Wet lab journal/2010/07/06

{| width="800"
 * style="background-color: #EEE"|[[Image:igem-logo-150px.png|150px]] WiFi coli: A communi-colight system
 * style="background-color: #F2F2F2" align="center"|  |Main project page
 * style="background-color: #F2F2F2" align="center"|  |Main project page


 * colspan="2"|
 * colspan="2"|

Back to the lab
      1. Ladder.  2. 1.2 p-s.  3. 1.7 p-s.  4. 1.9 p-s.  5. 2.2 p-s.  6. 2.8 p-s.  7. 3.1 p-s.  8. 3.3 p-s.  9. 3.5 p-s. <br/ > 10. 3.8 p-s. <br/ > 11. 3.9 p-s. <br/ > 12. 4.2 p-s. <br/ > 13. 4.3 p-s. <br/ > 14. Ladder. <br/ > 15. Empty. <br/ > 16. Ladder. <br/ > 17. 4.7 p-s. <br/ > 18. 4.8 p-s. <br/ > 19. 4.9 p-s. <br/ > 20. 2.2 <br/ > 21. 2.8 <br/ > 22. 4.2 <br/ > 23. 4.3 <br/ > 24. 4.7 <br/ > 25. 4.8 <br/ > 26. 4.9 <br/ > 27. Co Neg PCR<br/ > 28. Co Pos PCR (mut-luz PCR)<br/ > <br/ > <br/ >
 * Today, I repeated the gel that I couldn't see the other day.
 * The lanes were as follows:

Me against the gel
<br/ > <br/ > <br/ > <br/ > <br/ > 1. Ladder <br/ >
 * In order to quantify the ligation, I ran a gel for 40 min at 95V, which is observed next:
 * The lanes were as follows:

2. Dephosphated plasmid with J23106 <br/ >

3. Dephosphated plasmid 18 <br/ >

4. Restriction of Luciferase <br/ >

5. Restriction of Mutated luciferase <br/ >

<br/ >

Kind of getting used to ligations...
<br/ > <br/ > <br/ > -H2O ---> 9μL<br/ > -Buffer ligase ---> 2μL<br/ > -Vector (plasmid) -> 2μL<br/ > -Insert (luciferase) ---> 6μL<br/ > -Ligase > 1μL<br/ > <br/ > <br/ > -H2O ---> 8μL<br/ > -Buffer ligase ---> 2μL<br/ > -Vector (plasmid) -> 3μL<br/ > -Insert (luciferase) ---> 6μL<br/ > -Ligase > 1μL<br/ > <br/ > <br/ > -H2O ---> 10μL<br/ > -Buffer ligase ---> 2μL<br/ > -Vector (plasmid) -> 2μL<br/ > -Insert (luciferase) ---> 5μL<br/ > -Ligase > 1μL<br/ > <br/ > <br/ > -H2O ---> 9μL<br/ > -Buffer ligase ---> 2μL<br/ > -Vector (plasmid) -> 3μL<br/ > -Insert (luciferase) ---> 5μL<br/ > -Ligase > 1μL<br/ > <br/ > <br/ > -H2O ---> 15μL<br/ > -Buffer ligase ---> 2μL<br/ > -Vector (plasmid) -> 2μL<br/ > -Ligase > 1μL<br/ > <br/ > <br/ > -H2O ---> 14μL<br/ > -Buffer ligase ---> 2μL<br/ > -Vector (plasmid) -> 3μL<br/ > -Ligase > 1μL<br/ > <br/ > <br/ >
 * As I am planning on repeating the experiments, I did the ligations again...
 * 1: luciferase (green) with J23106
 * 2: luciferase (green) with plasmid 18
 * 3: luciferase (red) with J23106
 * 4: luciferase (red) with plasmid 18
 * 5: Control (J23106)
 * 6: Control (plasmid 18)
 * The reactions were left at 16°C ON.


 * }