Difference between revisions of "Team:BOKU-Vienna/Protocol"

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#interlab {
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<h4 id="transformation">Transformation of chemically competent E. Coli</h4>
 
<h4 id="transformation">Transformation of chemically competent E. Coli</h4>
 
<p></p>
 
<p></p>
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<p> Competent cells from E. Coli strain DH5α or DH10B were thawed on ice. The whole ligation/other DNA samples were added to the cells, for 30 min the mix was put on ice. The cells and DNA mix were heat shocked for 90s at 42°C. The cells recovered on ice for 5min. 1 mL of LB medium was added to the cells and then followed an incubation at 37°C.
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<p> Competent cells from E. Coli strain DH5α or DH10B were thawed on ice. The whole ligation/other DNA samples were added to the cells, for 30 min the mix was put on ice. The cells and DNA mix were heat shocked for 90s at 42°C. The cells recovered on ice for 5min. 1 mL of LB medium was added to the cells and then an incubation followed at 37°C in a shaker-incubator for 45min. An appropiate amount of cells were plated on a selective LB agar.</p>
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shaker-incubator (150 rpm) for 45 min to 1 h. Plate the appropriate amount of cells on selective LB agar.</p>
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<p>Minipreps were performed using Hi Yield®Plasmid Mini DNA Isolationkit according to the manufacturer's protocols.
 
<p>Minipreps were performed using Hi Yield®Plasmid Mini DNA Isolationkit according to the manufacturer's protocols.
 
</p>
 
</p>
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<h4 id="interlab">InterLab Challenge</h4>
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<p></p>
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<p>Interlab Challenge was performed according to the protocols for the InterLab Study in 2017. For further details visit the iGEM homepage: <a href="https://2017.igem.org/Competition/InterLab_Study">InterLab Study</a> or our <a href=" https://2017.igem.org/Team:BOKU-Vienna/InterLab">InterLab page</a>.</p>
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<h5>Kit Plate 6 InterLab Part Locations</h5>
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<table class="table">
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<thead>
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<tr>
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<th>Name</th>
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<th>Backbone</th>
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</tr>
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</thead>
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<tbody>
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<tr>
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<td>Positive Control</td>
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<td>BBa_I20270</td>
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</tr>
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<tr>
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<td>Negative Control</td>
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<td>BBa_R0040</td>
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</tr>
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        <tr>
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<td>Test Device 1</td>
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<td>BBa_J364000</td>
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</tr>
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        <tr>
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<td>Test Device 2</td>
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<td>BBa_J364001</td>
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</tr>
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        <tr>
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<td>Test Device 3</td>
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<td>BBa_J364002</td>
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</tr>
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        <tr>
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<td>Test Device 4/td>
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<td>BBa_J364003</td>
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</tr>
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        <tr>
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<td>Test Device 5</td>
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<td>BBa_J364004</td>
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</tr>
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        <tr>
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<td>Test Device 6</td>
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<td>BBa_J364005</td>
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</tr>
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</tbody>
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</table>
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<p></p>
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</div>
 
</div>
  

Revision as of 14:25, 10 July 2017

Protocol

V

Protocols from A-Z.


Here you will find our protocols in the future.

Contents

Materials

LB medium - E. Coli

Name Amount
Hy Soy Peptone 10.0g
Yeast Extract 5.0g
NaCl 5.0g
RO-water 1000 mL

LB Agar - E. Coli

Name Amount
Hy soy Peptone 10.0g
Yeast Extract 5.0g
NaCl 5.0g
RO-water 980 mL
Agar-Agar per 500mL 10g

Methods

Transformation of chemically competent E. Coli

Competent cells from E. Coli strain DH5α or DH10B were thawed on ice. The whole ligation/other DNA samples were added to the cells, for 30 min the mix was put on ice. The cells and DNA mix were heat shocked for 90s at 42°C. The cells recovered on ice for 5min. 1 mL of LB medium was added to the cells and then an incubation followed at 37°C in a shaker-incubator for 45min. An appropiate amount of cells were plated on a selective LB agar.

Miniprep

Minipreps were performed using Hi Yield®Plasmid Mini DNA Isolationkit according to the manufacturer's protocols.

InterLab Challenge

Interlab Challenge was performed according to the protocols for the InterLab Study in 2017. For further details visit the iGEM homepage: InterLab Study or our InterLab page.

Kit Plate 6 InterLab Part Locations
Name Backbone
Positive Control BBa_I20270
Negative Control BBa_R0040
Test Device 1 BBa_J364000
Test Device 2 BBa_J364001
Test Device 3 BBa_J364002
Test Device 4/td> BBa_J364003
Test Device 5 BBa_J364004
Test Device 6 BBa_J364005