Summary
Digest All the Things
Well One: Ladder
Well Two
| Reagent
|
Volume
|
Expected:
1. Unknown
Observed:
3k, 7k, 2k
|
| DNA(CMV/MV9) |
5.0 μL
|
| 10X buffer |
1.5 μL
|
| XbaI |
0.0 μL
|
| NruI |
0.0 μL
|
| dH2O |
8.5 μL
|
| Total |
15 μL --> 37°C/ 30 min.
|
Well Three
| Reagent
|
Volume
|
Expected:
1. 5200bp
Observed:
5.2k, 2k
|
| DNA(CMV/MV9) |
5.0 μL
|
| 10X buffer |
1.5 μL
|
| XbaI |
1.0 μL
|
| NruI |
0.0 μL
|
| dH2O |
7.5 μL
|
| Total |
15 μL --> 37°C/ 30 min.
|
Well Four
| Reagent
|
Volume
|
Expected:
1. 5200bp
Observed:
5.2k, 2k
|
| DNA(CMV/MV9) |
5.0 μL
|
| 10X buffer |
1.5 μL
|
| XbaI |
0.0 μL
|
| NruI |
1.0 μL
|
| dH2O |
7.5 μL
|
| Total |
15 μL --> 37°C/ 30 min.
|
Well Five
| Reagent
|
Volume
|
Expected:
1. 710bp, 4490bp
Observed:
4k, 2k, 700
|
| DNA(CMV/MV9) |
5.0 μL
|
| 10X buffer |
1.5 μL
|
| XbaI |
1.0 μL
|
| NruI |
1.0 μL
|
| dH2O |
6.5 μL
|
| Total |
15 μL --> 37°C/ 30 min.
|
Well Six
| Reagent
|
Volume
|
Expected:
1. BL01 = Insert (2520bp), V0120(3265bp)
Observed:
3.1k, 2.6k
|
| DNA(BL01A) |
5.0 μL
|
| 10X buffer |
1.5 μL
|
| XbaI |
1.0 μL
|
| SpeI |
1.0 μL
|
| dH2O |
7.5 μL
|
| Total |
15 μL --> 37°C/ 30 min.
|
Well Seven
| Reagent
|
Volume
|
Expected:
1. BL01 = 2520bp, 3265bp
Observed:
3.1k, 2.6k, 7k
|
| DNA(BL01B) |
5.0 μL
|
| 10X buffer |
1.5 μL
|
| XbaI |
1.0 μL
|
| SpeI |
1.0 μL
|
| dH2O |
7.5 μL
|
| Total |
15 μL --> 37°C/ 30 min.
|
Well Eight
| Reagent
|
Volume
|
Expected:
2520bp, 3265bp Observed:
Faint: 3.1k, 2.6k, 7k
|
| DNA(BL05) |
5.0 μL
|
| 10X buffer |
1.5 μL
|
| XbaI |
1.0 μL
|
| SpeI |
1.0 μL
|
| dH2O |
7.5 μL
|
| Total |
15 μL --> 37°C/ 30 min.
|
Well Nine
| Reagent
|
Volume
|
Expected:
1431bp, 3265bp Observed:
3.1k, 4.9k, 1.5k
|
| DNA(BL09) |
5.0 μL
|
| 10X buffer |
1.5 μL
|
| XbaI |
1.0 μL
|
| SpeI |
1.0 μL
|
| dH2O |
7.5 μL
|
| Total |
15 μL --> 37°C/ 30 min.
|
Well 10
| Reagent
|
Volume
|
Expected:
1. BL01 = 7720bp
Observed:
4k, 2k, 1.5k
|
| DNA(CMV/MV9 & BL01) |
5.0 μL
|
| 10X buffer |
1.5 μL
|
| XbaI |
0.0 μL
|
| NruI |
1.0 μL
|
| dH2O |
7.5 μL
|
| Total |
15 μL --> 37°C/ 30 min.
|
Well 11
| Reagent
|
Volume
|
Expected:
1. dependent on ligation direction
Observed:
4k, 2k, 1.5k
|
| DNA(CMV/MV9 & BL01) |
5.0 μL
|
| 10X buffer |
1.5 μL
|
| XbaI |
1.0 μL
|
| NruI |
1.0 μL
|
| dH2O |
6.5 μL
|
| Total |
15 μL --> 37°C/ 30 min.
|
Gel Verification
|
