green light receptor
Order primers
Investigators: Jakob
- Order new primers for the quickchange of CcaS and CcaR
blue light receptor
Transformation
Investigators: Sophie
Transformation of ligated parts:
- LovTAP
- Not-Gate
- T3 vector
stored in incubator at 37°C.
red light receptor
PCR for Cph8
Investigators:Julia
after protocol from Uppsala.
PCR-Mixture for one Reaction:
For a 50 µl reaction use
| 32,5µl
| H20
|
|
| 10µl
| 5x Phusion Buffer
|
|
| 2.5µl
| Primer fw
|
|
| 2.5µl
| Primer dw
|
|
| 1µl
| dNTPs
|
|
| 1µl
| DNA-Template
|
|
| 0.5 µl
| Phusion (add in the end)
|
|
Primer used:
cph8 Prefix: TTCGAATTCGCGGCCGCTTCTAGATGGCCACCACCGTACAA
cph8 Suffix: CCGCTACTAGTATTATTACCCTTCTTTTGTCATGCCCT
PCR program:
Temp. Time,Nr of cycles
98° 5' 1x
98° 30 10x
65° 30
72° 1'15
98° 30 20x
72° 1'15
72° 7' 1x
4° ∞
Template: Voights plasmid (10x dilution)
Amplicon: 2235 bp
Elongation time Phusion 67,05 sec
Lysis cassette
NAME OF YOUR EXPERIMENT
Investigators:NAME
Precipitator
Ligation
Investigators: Sophie
continue from experiment: Cloning (12.8.11)
Project name: inducible promoter for pbd
Vector: psb1T3
Inserts: IPTG-Promoter,Pbd-GFP
samples stored in Ruediger's box
NAME OF YOUR EXPERIMENT: Transformation
Investigators: Sophie
continue from experiment: Ligation
Project name: inducible promoter for pbd
samples stored in incubator
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