Team:Groningen/Labwork/8 August 2013

From 2013.igem.org

(Difference between revisions)
(Created page with "<html> <head> <meta http-equiv="X-UA-Compatible" content="IE=EDGE" charset="utf-8" /> <link rel="stylesheet" href="https://2013.igem.org/Team:Groningen/CSS/DefaultPage?action=ra...")
 
(6 intermediate revisions not shown)
Line 26: Line 26:
<br>Inoculated the colony of CFP and four colonies of Pdes/CheY/eYFP in LB medium with amp for mini prep.  
<br>Inoculated the colony of CFP and four colonies of Pdes/CheY/eYFP in LB medium with amp for mini prep.  
<br>Made a new ligation reaction between Pdes and CheY.  
<br>Made a new ligation reaction between Pdes and CheY.  
 +
<br>Made a new ligation reaction between CFP and the backbone.
 +
<br>Made a new ligation reaction between Pdes/CheY and the backbone.
 +
<br>Transformation to <i>E. coli</i> of all the ligation reactions.
 +
<h2>Claudio</h2>
 +
Yesterday 2x SG plates in the presence of titanium are prepared.
 +
<br>2&micro;l of <i>Bacillus Subtilis</i> overnight culture is spotted on titanium in the presence of 2x SG and without. The plates are incubated at 30&deg;C.
 +
<br>Two of the <i>Bacillus Subtilis</i> complex colonies, which grew during the last few days on 2x SG, were transferred onto titanium in the presence of 2x SG and without. The plates are incubated at 30&deg;C.
 +
<br>100&micro;l of the <i>Bacillus Subtilis</i> overnight culture was inoculated in 20 ml of SGG medium. The bottle is incubated at 30&deg;C.
 +
<br>Pictures of the biofilm are taken over time.
 +
 +
<h2>Sebas</h2>
 +
there were approx. 100 colonies per plate, however a vague smear was visibile on the negative control. Restreaked 6 <br>colonies per plate on new LB plates(5ug/ml cam).
 +
<br>
 +
<br>Did prep on 4 colonies harboring the potential Pdes-cheY-eyfp plasmid. Did a digestion with EcoRI*PstI.
 +
<br>Expected sizes: 6064 and 1409, 
 +
<br>sizes on gel: 6000 and 1100.
 +
<br>Indicating that a part of 300bp is missing (probably the Pdes part)
</div>
</div>

Latest revision as of 17:19, 8 August 2013

Mirjam

Check of the plates of the restrictions made yesterday. One of the plates with CFP showed 1 colony. The other two plates (one IPTG) did not show any. The transformation of Pdes/CheY into the backbone failed, but the transformation of Pdes/CheY/eYFP showed a couple of colonies.
Inoculated the colony of CFP and four colonies of Pdes/CheY/eYFP in LB medium with amp for mini prep.
Made a new ligation reaction between Pdes and CheY.
Made a new ligation reaction between CFP and the backbone.
Made a new ligation reaction between Pdes/CheY and the backbone.
Transformation to E. coli of all the ligation reactions.

Claudio

Yesterday 2x SG plates in the presence of titanium are prepared.
2µl of Bacillus Subtilis overnight culture is spotted on titanium in the presence of 2x SG and without. The plates are incubated at 30°C.
Two of the Bacillus Subtilis complex colonies, which grew during the last few days on 2x SG, were transferred onto titanium in the presence of 2x SG and without. The plates are incubated at 30°C.
100µl of the Bacillus Subtilis overnight culture was inoculated in 20 ml of SGG medium. The bottle is incubated at 30°C.
Pictures of the biofilm are taken over time.

Sebas

there were approx. 100 colonies per plate, however a vague smear was visibile on the negative control. Restreaked 6
colonies per plate on new LB plates(5ug/ml cam).

Did prep on 4 colonies harboring the potential Pdes-cheY-eyfp plasmid. Did a digestion with EcoRI*PstI.
Expected sizes: 6064 and 1409,
sizes on gel: 6000 and 1100.
Indicating that a part of 300bp is missing (probably the Pdes part)