Back to the lab
- Today, I repeated the gel that I couldn't see the other day.
- The lanes were as follows:
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.
10. 3.8 p-s.
11. 3.9 p-s.
12. 4.2 p-s.
13. 4.3 p-s.
14. Ladder.
15. Empty.
16. Ladder.
17. 4.7 p-s.
18. 4.8 p-s.
19. 4.9 p-s.
20. 2.2
21. 2.8
22. 4.2
23. 4.3
24. 4.7
25. 4.8
26. 4.9
27. Co Neg PCR
28. Co Pos PCR (mut-luz PCR)
Me against the gel
- In order to quantify the ligation, I ran a gel for 40 min at 95V, which is observed next:
- The lanes were as follows:
1. Ladder
2. Dephosphated plasmid with J23106
3. Dephosphated plasmid 18
4. Restriction of Luciferase
5. Restriction of Mutated luciferase
Kind of getting used to ligations...
- As I am planning on repeating the experiments, I did the ligations again...
- 1: luciferase (green) with J23106
-H2O -----------------------> 9μL
-Buffer ligase -----------> 2μL
-Vector (plasmid) -----> 2μL
-Insert (luciferase) ---> 6μL
-Ligase --------------------> 1μL
- 2: luciferase (green) with plasmid 18
-H2O -----------------------> 8μL
-Buffer ligase -----------> 2μL
-Vector (plasmid) -----> 3μL
-Insert (luciferase) ---> 6μL
-Ligase --------------------> 1μL
- 3: luciferase (red) with J23106
-H2O -----------------------> 10μL
-Buffer ligase -----------> 2μL
-Vector (plasmid) -----> 2μL
-Insert (luciferase) ---> 5μL
-Ligase --------------------> 1μL
- 4: luciferase (red) with plasmid 18
-H2O -----------------------> 9μL
-Buffer ligase -----------> 2μL
-Vector (plasmid) -----> 3μL
-Insert (luciferase) ---> 5μL
-Ligase --------------------> 1μL
-H2O -----------------------> 15μL
-Buffer ligase -----------> 2μL
-Vector (plasmid) -----> 2μL
-Ligase --------------------> 1μL
-H2O -----------------------> 14μL
-Buffer ligase -----------> 2μL
-Vector (plasmid) -----> 3μL
-Ligase --------------------> 1μL
- The reactions were left at 16°C ON.
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