Team:Groningen/Labwork/17 July 2013

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'''Mirjam'''
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<br/>Restriction digest of the 1st and 3rd plate colonies are examined on gel. 5/7 colonies are correct transformations. To be completely sure if the transformation did work, a PCR for the promoter is made, using the colonies and the original construct as a backbone.  
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<br/>Miniprep for the samples of the 4th plate (they grew O/N). Restriction analysis with EcoRV.
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<br/>Run gel of the restriction analysis.
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<h2>Mirjam</h2>
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<br/>Check of the restriction done yesterday. This revealed that 5/7 colonies did contain the transformant. The cells of the fourth plate started to grow, so a mini prep analysis is performed. A <a href="https://2013.igem.org/Team:Groningen/protocols/Digestion"><FONT COLOR="black"><b>restriction analysis</b></FONT></a> is done with EcoRV.
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A second confirmation of the constructs is done using <a href="https://2013.igem.org/Team:Groningen/protocols/PCR"><FONT COLOR="black"><b>PCR</b></FONT></a>. <a href="https://2013.igem.org/Team:Groningen/protocols/GelElectrophoresis"><FONT COLOR="black"><b>Gel electrophorese</b></FONT></a>  analysis revealed that indeed the colonies contain the promoter, although one of them only in small amount. So it is chosen to remove that one from the stock.
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Miniprep is done to obtain more biobrick BBa_K823023.
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YFP, RFP and GFP are searched in the biobrick system and found to be in the spring 2013 plates. A transformation to <i>E. coli</i> is done for these biobricks. Along with a new <a href="https://2013.igem.org/Team:Groningen/protocols/Transformation_EC"><FONT COLOR="black"><b>transformation</b></FONT></a> of one of the transformants (to do some IPTG tests with).
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<h2>Inne en Sander</h2>
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<br>The <i>E. coli</i> cells with the BBa_k818000 that were to grow overnight didn't grow.
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<br>The tubes that were supposed to be turbid were still clear, however the negative control was also clear so that one did succeed.
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<br> Made -80 C stock of the Munich backbone BBa_K823023, in tray iGEM 2012 A, position 26.
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<br> Inoculated the BBa_818000, made 5 tubes. Inoculation takes place O/N at 37 C.
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<td>Sample</td>
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<td>Composition</td>
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<td>1</td>
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<td>4mL LB + 8uL Amp</td>
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<td>2</td>
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<td>4mL LB + 8uL Amp</td>
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<td>3</td>
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<td>4mL LB + 8uL Amp (negative control)</td>
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<td>4</td>
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<td>4mL LB</td>
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<td>5</td>
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<td>4mL LB (negative control)</td>
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<h2>Claudio and Mike</h2>
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<br>The silk synthetic genes design is continued: all the different constructs which are necessary in order to get the highest chance of success are produced.
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  <p>iGEM 2013 Groningen</p>
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Latest revision as of 10:38, 30 July 2013

Mirjam


Check of the restriction done yesterday. This revealed that 5/7 colonies did contain the transformant. The cells of the fourth plate started to grow, so a mini prep analysis is performed. A restriction analysis is done with EcoRV. A second confirmation of the constructs is done using PCR. Gel electrophorese analysis revealed that indeed the colonies contain the promoter, although one of them only in small amount. So it is chosen to remove that one from the stock. Miniprep is done to obtain more biobrick BBa_K823023. YFP, RFP and GFP are searched in the biobrick system and found to be in the spring 2013 plates. A transformation to E. coli is done for these biobricks. Along with a new transformation of one of the transformants (to do some IPTG tests with).

Inne en Sander


The E. coli cells with the BBa_k818000 that were to grow overnight didn't grow.
The tubes that were supposed to be turbid were still clear, however the negative control was also clear so that one did succeed.
Made -80 C stock of the Munich backbone BBa_K823023, in tray iGEM 2012 A, position 26.
Inoculated the BBa_818000, made 5 tubes. Inoculation takes place O/N at 37 C.
Sample Composition
1 4mL LB + 8uL Amp
2 4mL LB + 8uL Amp
3 4mL LB + 8uL Amp (negative control)
4 4mL LB
5 4mL LB (negative control)

Claudio and Mike


The silk synthetic genes design is continued: all the different constructs which are necessary in order to get the highest chance of success are produced.

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