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EPF-Lausanne/8 September 2009
From 2009.igem.org
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==Wet Lab== | ==Wet Lab== | ||
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===Cultures=== | ===Cultures=== | ||
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'''Results:''' | '''Results:''' | ||
:RO1 has grown in LB and M9 | :RO1 has grown in LB and M9 | ||
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:RO2+BB #1,4 did not grow | :RO2+BB #1,4 did not grow | ||
:RO2+BB #3 has grown in LB but not in M9 | :RO2+BB #3 has grown in LB but not in M9 | ||
| - | -> problem with our glycerol stock. We did some plates of the cultures that grew. | + | -> problem with our glycerol stock (RO2 + BB didn't grow in LB !!). We did some plates of the cultures that grew. |
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| + | For safety we did some plates with the glycerol stocks of RO2 + BB 1,4,8. We also did some plates with the cultures that grew (RO1 1,2,3, RO2 4,5,10, RO2 + BB 3). | ||
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<br> | <br> | ||
===Characterization=== | ===Characterization=== | ||
As RO1 grew in both media, we put it in 25mL medium with Amp. We did also with RO2 (LB), and RO2+BB (LB), and LacI-RFP. | As RO1 grew in both media, we put it in 25mL medium with Amp. We did also with RO2 (LB), and RO2+BB (LB), and LacI-RFP. | ||
| + | For the characterisation, we did different combinations : -TRP, -TRP -ATC, +TRP -ATC, +TRP +ATC. Evertime for all the clones in M9 and then in LB. | ||
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| + | TRP : LB : dilution 50x so 1ul, M9 : dilution 25x so 2 ul. We took the stock solution 4,250 g/L. | ||
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| + | ATC : 5x dilution so 10 ul from the 500 ng/ml stock solution. | ||
Revision as of 09:30, 21 September 2009
Wet Lab
Cultures
Results:
- RO1 has grown in LB and M9
- RO2 has grown in LB but not in M9
- RO2+BB #1,4 did not grow
- RO2+BB #3 has grown in LB but not in M9
-> problem with our glycerol stock (RO2 + BB didn't grow in LB !!). We did some plates of the cultures that grew.
For safety we did some plates with the glycerol stocks of RO2 + BB 1,4,8. We also did some plates with the cultures that grew (RO1 1,2,3, RO2 4,5,10, RO2 + BB 3).
Characterization
As RO1 grew in both media, we put it in 25mL medium with Amp. We did also with RO2 (LB), and RO2+BB (LB), and LacI-RFP.
For the characterisation, we did different combinations : -TRP, -TRP -ATC, +TRP -ATC, +TRP +ATC. Evertime for all the clones in M9 and then in LB.
TRP : LB : dilution 50x so 1ul, M9 : dilution 25x so 2 ul. We took the stock solution 4,250 g/L.
ATC : 5x dilution so 10 ul from the 500 ng/ml stock solution.
to be completed ...
People in the lab
Mélanie, Caroline, Basile, Nicolas
