green light receptor
NAME OF YOUR EXPERIMENT
Investigators:NAME
blue light receptor
Gel purification
Investigators:Sophie
the bands of LOV-pcr and NOT-pcr where purificated. The vector was digested again because the band on the gel was not intense enough.
Ligation
Investigators: Sophie:
LOV-pcr, NOT-pcr and the Amp vector where ligated at 16°C
red light receptor
PCR
Investigator: Jakob
PCR
Name:
Jakob
| Date:
16.08.2011
|
Continue from Experiment: 12.08.2011
Order primers
|
Project Name:
Red light receptor, amplify Cph8
Quickchange of CcaR (EcoRI)
|
PCR-Mixture for one Reaction:
For a 50 µl reaction use
32,5µl
| H20
| Name
|
10µl
| 5x Phusion Buffer
| of Primer
|
2.5µl
| Primer fw
| P 58 (1:10)
|
2.5µl
| Primer dw
| P 59 (1:10)
|
1µl
| dNTPs
| of Template DNA
|
1µl
| DNA-Template
| JT122 5ng/µl
|
0.5 µl
| Phusion (add in the end)
|
|
The program we used: for Quickchange of CcaR (EcoRI)
Temp.
| Time
| Nr. of cycles
|
94°
| 5’
| 1x
|
94°
| 30’’
| 2x
|
64°
| 30’’
|
72°
| 1’30’’
|
94°
| 30’’
| 2x
|
62°
| 30’’
|
72°
| 1’30’’
|
94°
| 30’’
| 5x
|
61,6°
| 30’’
|
72°
| 1’30’’
|
94°
| 30’’
| 25x
|
57,8°
| 30’’
|
72°
| 1’30’’
|
72°
| 7’
| 1x
|
4°
| ∞
|
Lysis cassette
NAME OF YOUR EXPERIMENT
Investigators:NAME
Precipitator
NAME OF YOUR EXPERIMENT
Investigators: NAME