Team:Groningen/Labwork/12 August 2013

From 2013.igem.org

(Difference between revisions)
Line 23: Line 23:
<h1>Sander</h1>
<h1>Sander</h1>
-
did a miniprep of CFP 1,2 and 4 and Pdes CheY 1 and 2 and a gell purification of Munich EXP
+
did a miniprep of CFP 1,2 and 4 and Pdes CheY 1 and 2 and a gell purification of Munich EcoRI*PstI
<h2>Inne</h2>
<h2>Inne</h2>
Line 33: Line 33:
And a cone (salvaged from a 5 ml pipet) seperating the strain from air.
And a cone (salvaged from a 5 ml pipet) seperating the strain from air.
<br>Tubes are to grow overnight in a 37C incubator. Except one of the duplo series which was put in the shaker to see the difference.
<br>Tubes are to grow overnight in a 37C incubator. Except one of the duplo series which was put in the shaker to see the difference.
 +
 +
<h2>Sebas</h2>
 +
<br>Inocculated GFP 1&2 YFP 1&2 1:50 in 20 ml of LB.
 +
<br>At OD600=~0,4 took sample T0 and added 1mM of IPTG
 +
<br>Grew for 2 hours and checked under the microscope.
 +
<br>YFP1 did not give any signal.
 +
<br>GFP1 gave a weak signal for T0&T2
 +
<br>
 +
<br>Need to do a better setup with B. subtilis 168 wt as negative control.
 +
<br>so, inoccualted GFP 1&2 and B.sub168 wt.
 +
<Br>
 +
<br><b>Discussion:</b><br> GFP was token from the BioBrick system (Claudio, add link please) and is not optimized for B. subtilis. eYFP (Mirjam add link please) didn't had a RBS and <br>this may cause no translation of the protein.
</div>
</div>

Revision as of 20:05, 12 August 2013

Sander

did a miniprep of CFP 1,2 and 4 and Pdes CheY 1 and 2 and a gell purification of Munich EcoRI*PstI

Inne

Made -80 stock of strains OI3017 OI3059 OI2653 OI3135.
Made a setup to test motility, using 0.4% agar test-tubes with different obstacles in them.
4 sets of 3 tubes are stabbed with a toothpick, one negative, one with Bacillus subtillus gfp backbone, and one OI3017.
The OI3017 should not be motile the Bacillus subtillus gfp should be motile and the negative control should be negative.
The normal test was done in duplo (due to a failed abstacle preparation). the other test are 3 ml agar on top of the normal level. And a cone (salvaged from a 5 ml pipet) seperating the strain from air.
Tubes are to grow overnight in a 37C incubator. Except one of the duplo series which was put in the shaker to see the difference.

Sebas


Inocculated GFP 1&2 YFP 1&2 1:50 in 20 ml of LB.
At OD600=~0,4 took sample T0 and added 1mM of IPTG
Grew for 2 hours and checked under the microscope.
YFP1 did not give any signal.
GFP1 gave a weak signal for T0&T2

Need to do a better setup with B. subtilis 168 wt as negative control.
so, inoccualted GFP 1&2 and B.sub168 wt.

Discussion:
GFP was token from the BioBrick system (Claudio, add link please) and is not optimized for B. subtilis. eYFP (Mirjam add link please) didn't had a RBS and
this may cause no translation of the protein.