Team:Groningen/Labwork/9 August 2013

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Monitoring the grow of biofilm in liquid medium, LB agar and on titanium only.
Monitoring the grow of biofilm in liquid medium, LB agar and on titanium only.
 +
<h2>Sebas</h2>
 +
2 colonies on Pdes::cheY in munich backbone plate, did colony pcr the expected 675bp band was visible with some <br>aspecific bands. Inoculated both colonies in 3ml/LB/amp o/n at 37degrees celcius.
 +
<br>There were approx 150 colonies on the Munich+hyperspank+CFP plate. Inoculated 4col. in 3ml/LB/amp o/n 37degerees.
 +
<br>There were approx 100 colonies on the Pdes::cheY:eYFP in Munich backbone plate. All colonies were red indicating <br>that the P::RFP was still there.
 +
<br>Did a digestion on munich backbone with pstI and EcoRI with 1ug DNA for 30min at 37degrees celcius.
 +
<br>cut out the backbone from gel and stored at -20 degrees celcius
 +
<br>The 6 colonies for GFP and YFP in b.sub grew on new plates -> stabbed colonies on starch plate and incubated o/n <br>at 37degees celcius.
 +
 +
<h2>Inne & Sander</h2>
 +
Received the motility knock outs today. Plated them on plates with various selection markers.
 +
<table border="1">
 +
<tr>
 +
<td>strain</td>
 +
<td>description</td>
 +
<td>selection marker</td>
 +
</tr>
 +
<tr>
 +
<td>OI2838</td>
 +
<td>cheB::cat</td>
 +
<td>5 uL/mL cm</td>
 +
</tr>
 +
<tr>
 +
<td>OI3305</td>
 +
<td>cheC1 cheD1::cat</td>
 +
<td>5 uL/mL cm</td>
 +
</tr>
 +
<tr>
 +
<td>OI2737</td>
 +
<td>cheW::cat</td>
 +
<td>5 uL/mL cm</td>
 +
</tr>
 +
<tr>
 +
<td>OI3017</td>
 +
<td>cheB/cheR::cat</td>
 +
<td>5 uL/mL cm</td>
 +
</tr>
 +
<tr>
 +
<td>OI2653</td>
 +
<td>delta cheR</td>
 +
<td>5 uL/mL cm</td>
 +
</tr>
 +
<tr>
 +
<td>OI2653</td>
 +
<td>delta cheR</td>
 +
<td>none</td>
 +
</tr>
 +
<tr>
 +
<td>OI3305</td>
 +
<td>delta cheC1</td>
 +
<td>none</td>
 +
</tr>
 +
<tr>
 +
<td>OI3061</td>
 +
<td>cheW::cat,cheV::kan</td>
 +
<td>5 uL/mL cm 10 uL/mL kan</td>
 +
</tr>
 +
<td>OI3059</td>
 +
<td>cheV::kan</td>
 +
<td>10 uL/mL kan</td>
 +
</tr>
 +
</table>
</div>
</div>

Latest revision as of 19:03, 9 August 2013

Claudio and Sander

Monitoring the grow of biofilm in liquid medium, LB agar and on titanium only.

Sebas

2 colonies on Pdes::cheY in munich backbone plate, did colony pcr the expected 675bp band was visible with some
aspecific bands. Inoculated both colonies in 3ml/LB/amp o/n at 37degrees celcius.
There were approx 150 colonies on the Munich+hyperspank+CFP plate. Inoculated 4col. in 3ml/LB/amp o/n 37degerees.
There were approx 100 colonies on the Pdes::cheY:eYFP in Munich backbone plate. All colonies were red indicating
that the P::RFP was still there.
Did a digestion on munich backbone with pstI and EcoRI with 1ug DNA for 30min at 37degrees celcius.
cut out the backbone from gel and stored at -20 degrees celcius
The 6 colonies for GFP and YFP in b.sub grew on new plates -> stabbed colonies on starch plate and incubated o/n
at 37degees celcius.

Inne & Sander

Received the motility knock outs today. Plated them on plates with various selection markers.
strain description selection marker
OI2838 cheB::cat 5 uL/mL cm
OI3305 cheC1 cheD1::cat 5 uL/mL cm
OI2737 cheW::cat 5 uL/mL cm
OI3017 cheB/cheR::cat 5 uL/mL cm
OI2653 delta cheR 5 uL/mL cm
OI2653 delta cheR none
OI3305 delta cheC1 none
OI3061 cheW::cat,cheV::kan 5 uL/mL cm 10 uL/mL kan
OI3059 cheV::kan 10 uL/mL kan