Goals
- PCR for TerB0015-Backbone and PCR for HydEF1.
PCRs
PCR for TerB0015-Backbone
Reagents
|
μL
|
MegaMix
|
H2O
|
24.5
|
50
|
5X Phusion Buffer HF
|
10
|
20.1
|
10mM dNTPs
|
1 each
|
2 each
|
Primer F
|
5
|
10.2
|
Primer R
|
5
|
10.2
|
DNA
|
1
|
2
|
Phusion
|
.5
|
NA
|
PCR for TerB0015-Backbone
Reagents
|
μL
|
MegaMix
|
H2O
|
23
|
46.5
|
5X Phusion Buffer HF
|
10
|
20.1
|
10mM dNTPs
|
1 each
|
2 each
|
Primer F
|
5
|
NA
|
Primer R
|
5
|
NA
|
DNA
|
1
|
2
|
DMSO
|
1.5
|
3
|
Phusion
|
.5
|
NA
|
Cycles
|
Gel instruct.
|
Gel
|
Terminators
- 98°C 30s
- 98°C 10s 30Cy
- 60°C 30S 30Cy
- 72°C 1:45 30Cy
- 72°C 10min
- 4°C
HydEF1
- 98°C 30s
- 98°C 10s 30Cy
- 60°C 30S 30Cy
- 72°C 50 30Cy
- 72°C 10min
- 4°C
|
- Lane1 Ladder
- Lane2 Ter1-Backbone
- Lane3 Ter2-Backbone
- Lane4 HydEF 1
- Lane5 HydEF 2
|
|
PCRs tube named as 1.0, 2.0, 3.0 and 4.0
|