Commons
Ligation
Investigators: Sandra
Ligation of PR1-PR6 with RFP (S1b and S2a). PR1-PR6 were digested with antarctica before start of ligation.
Start of ligation at 12:40. Incubation at room temperature for at least 3 hours.
Trafo
Investigators: Sandra
Trafo of:
- PR1+GFP
- PR2+GFP
- PR3+GFP
- PR4+GFP
- PR5+GFP
- PR6+GFP
green light receptor
NAME OF YOUR EXPERIMENT
Investigators:NAME
blue light receptor
no colonies on the plates
There were no colonies on the plates with ligated parts of the 2A-assembly.
Miniprep
Investigators: Sophie
yesterday two tubes with LB were inoculated with cells from our S35 glycerol stock(BBa_K322999). Today minipreps were performed.
PCR
Investigators: Sophie
| Name: Sophie
| Date: 07.09.11
|
| Continue from Experiment (Date)
(Name)
|
| Project Name: Gibson of ♥ and NOT
|
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
| ♥: P97
NOT: P99
|
| 2.5µl
| Primer dw
| ♥: P98
NOT: P100
|
| 1µl
| dNTPs
| of Template DNA
|
| 1µl
| DNA-Template
| M45a,b,c (BBa_Q0400)
Miniprep of M35 (1x) and M35b (2x) (BBa_K322999)
|
| 0.5 µl
| Phusion (add in the end)
|
|
Digestion with Dpn I and purification with PCR purification kit
What program do you use?
First 20 cycles touchdown 69°C -0.4/cycle
next 10 cycles touchdown 72°C -0.2 per cycle
PCR products were loaded onto a gel
red light receptor
NAME OF YOUR EXPERIMENT
Investigators:NAME
Lysis cassette
NAME OF YOUR EXPERIMENT
Investigators:NAME
Precipitator
Trafo
Investigators: Sandra
Trafo of the already ligated parts of the pbd + Gst-vector
Ligation
Investigators: Sophie
For pbd in iGEM C3-vector: again Ligation (see 31.08.11) because I don't find the last ligation product that produced positive clones. I have to redo the Trafo because the last positive clones seem to have mutated over the last generations, probably because the pbd is toxic.
labelled: L S54-ε5-C3 1:1 / 1:3
stored in "Ligationsreaktionen"-box
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