EPF-Lausanne/8 September 2009

From 2009.igem.org

(Difference between revisions)
(Wet Lab)
(Wet Lab)
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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.
 +
 
 +
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).
 +
 
<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.
 +
 +
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.

Revision as of 09:30, 21 September 2009

Contents

8 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