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Team:Freiburg/Notebook/7 September
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==<span style="color:grey;">Precipitator</span>== | ==<span style="color:grey;">Precipitator</span>== | ||
Revision as of 18:41, 7 September 2011
Contact 
Contents |
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
- PR1+50b
- PR2+50b
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
Troubleshooting of the modified Lysis genes K124017
Investigators:Theo
S4+S15 Nr1 and 7 were digested with XbaI and PstI, loaded on a 1% agarose gel and extracted on the 6th of September.
Today I digested the quickchanged-temperature sensitive promotor (K098995) with SpeI and PstI for ca 3 hours, incubated it with antarctic phosphatase for 1 hour and PCR-purified it.
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
