Difference between revisions of "Template:Team:Groningen/CONTENT/LOGBOOK/tasA Colony PCR t74"

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{{Team:Groningen/TEMPLATES/LOG
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{{Team:Groningen/TEMPLATES/EXPERIMENT
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|title=00:00, 18 August 2015 - 00:00, 18 August 2015
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|title=Experiment Title
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|executor=Harm Ruesink
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|protocol=Finibus sapien, nec cursus...
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|content=<div class="text">A new colony PCR was performed with 8 colonies from the T2+PliaG plate and control (T2). Each colony was resuspended in 20 µL MQ water. A Mastermix was made for 9 samples.</div>
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<div class="object data" id="tbl1">
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|description=Rutrum nulla. Nunc...
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<div class="wrapper">
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|conclusion=Malesuada a nisi ac...
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<div class="header">
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<div class="field fw3">Component</div>
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<div class="field fw7">Amount</div>
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</div>
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<div class="record">
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<div class="field fw3">Phusion polymerase</div>
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<div class="field fw7">2 µL</div>
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</div>
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<div class="record">
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<div class="field fw3">5x phusion buffer</div>
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<div class="field fw7">36 µL</div>
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</div>
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<div class="record">
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<div class="field fw3">DNTP MM (2 mM)</div>
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<div class="field fw7">18 µL</div>
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</div>
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<div class="record">
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<div class="field fw3">\( \mathrm{H_2O}\)</div>
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<div class="field fw7">120 µL</div>
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</div>
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<div class="record">
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<div class="field fw3">Primers (pSB1C3 colony PCR)</div>
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<div class="field fw7">3 µL each</div>
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</div>
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</div>
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<div class="caption">Components Mastermix.</div>
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</div>
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<div class="text">For the PCR samples, 18 µL of Mastermix was added to 4 µL of colony suspension. The following PCR thermocycle was used.</div>
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<div class="object data" id="tbl1">
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<div class="wrapper">
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<div class="header">
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<div class="field fw1">#</div>
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<div class="field fw3">Step</div>
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<div class="field fw2">Temperature</div>
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<div class="field fw3">Time</div>
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</div>
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<div class="record">
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<div class="field fw1">1</div>
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<div class="field fw3">Initial denaturation</div>
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<div class="field fw2">98 °C</div>
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<div class="field fw3">10:00</div>
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</div>
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<div class="record">
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<div class="field fw1">2</div>
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<div class="field fw3">Denaturation</div>
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<div class="field fw2">95 °C</div>
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<div class="field fw3">0:30</div>
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</div>
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<div class="record">
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<div class="field fw1">3</div>
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<div class="field fw3">Annealing</div>
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<div class="field fw2">55 °C</div>
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<div class="field fw3">0:30</div>
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</div>
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<div class="record">
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<div class="field fw1">4</div>
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<div class="field fw3">Extension</div>
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<div class="field fw2">72 °C</div>
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<div class="field fw3">1:30</div>
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</div>
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<div class="record">
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<div class="field fw1">5</div>
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<div class="field fw3">Go back to step 2 (repeat 30x)</div>
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<div class="field fw2"></div>
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<div class="field fw3"></div>
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</div>
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<div class="record">
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<div class="field fw1">6</div>
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<div class="field fw3">Final extension</div>
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<div class="field fw2">72 °C</div>
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<div class="field fw3">10:00</div>
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</div>
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</div>
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<div class="caption">PCR thermocycle.</div>
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</div>
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<div class="text">The samples were loaded on a 1% agarose gel with DNA stain G 1:50000. On the gel it was seen that the insert did not have the correct size, meaning that the promoter (PliaG) did not ligate in T2.</div>
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Revision as of 20:42, 18 September 2015

Experiment Title
Rutrum nulla. Nunc...
Ugue nulla, gravida non ante vel, hendrerit...
Malesuada a nisi ac...

<a class="postscriptum protocol" href="https://2015.igem.org/Team:Groningen/Protocols_and_Protocols/Finibus_sapien_nec_cursus...">Finibus sapien, nec cursus...</a>

Template:Team:Groningen/CONTENT/EXPERIMENTS/Experiment Title