10.3.10
10:30
diluted minipreps from yesterday to 20 ng/μL for PCR
PCR
Ingredient |
Amount |
Mastermix (10 reactions) |
10X Buffer |
2 μL |
20 μL |
10 mM dNTPs |
.4 μL |
4 μL |
Template |
20 ng (1 μL) |
added individually to each reaction |
F Primer |
.4 μL |
4 μL |
R Primer |
.4 μL |
4 μL |
Polymerase |
.2 μL |
2 μL |
H2O |
15.6 μL |
156 μL |
Samples
- J2334A215C 1
- J2334A215C 2
- J2334A215C 3
- J2334A215C 4
- ArsRSDM2C 1
- ArsRSDM2C 2
- ArsRSDM2C 3
- (+) control ArsR
- (-) control H2O
Primers
- VF2
- VR
- ArsRF EcoRI
- ArsR R
Program
95°C |
3 min |
95°C |
30 |
55°C |
30 sec |
72°C |
90 sec |
go to step 2 |
30X |
72°C |
3 min |
hold at 4°C |
Gel:
TAE 1% 175V
Lane |
Sample |
Amount |
1 |
1 Kb Ladder |
1 μL |
2 |
J2334A215 1 |
5 μL 1 μL dye |
3 |
J2334A215 2 |
5 μL 1 μL dye |
4 |
J2334A215 3 |
5 μL 1 μL dye |
5 |
J2334A215 4 |
5 μL 1 μL dye |
6 |
ArsRSDM2 1 |
5 μL 1 μL dye |
7 |
ArsRSDM2 2 |
5 μL 1 μL dye |
8 |
ArsRSDM2 3 |
5 μL 1 μL dye |
9 |
(+) control ArsR |
5 μL 1 μL dye |
10 |
(-) control |
5 μL 1 μL dye |
|