http://2012.igem.org/wiki/index.php?title=Team:Grenoble/Biology/Protocols/PCR_1&feed=atom&action=historyTeam:Grenoble/Biology/Protocols/PCR 1 - Revision history2024-03-28T12:05:46ZRevision history for this page on the wikiMediaWiki 1.16.0http://2012.igem.org/wiki/index.php?title=Team:Grenoble/Biology/Protocols/PCR_1&diff=166494&oldid=prevAdeline millet at 08:18, 25 September 20122012-09-25T08:18:07Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <tr><th><i><div class="petit"><center>SAFETY AND USEFUL RECOMMANDATION</center></div></i></th></tr></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <tr><th><i><div class="petit"><center>SAFETY AND USEFUL RECOMMANDATION</center></div></i></th></tr></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <tr><td><div class="petit">Those products are not dangerous at first sight, but in case of spreading</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <tr><td><div class="petit">Those products are not dangerous at first sight, but in case of spreading</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <tr><th><i><div class="petit"><center>SAFETY AND USEFUL RECOMMANDATION</center></div></i></th></tr></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <tr><th><i><div class="petit"><center>SAFETY AND USEFUL RECOMMANDATION</center></div></i></th></tr></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <tr><td><div class="petit">It was noticed that it exists a certain number of different thermocycler models.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <tr><td><div class="petit">It was noticed that it exists a certain number of different thermocycler models.</div></td></tr>
</table>Adeline millethttp://2012.igem.org/wiki/index.php?title=Team:Grenoble/Biology/Protocols/PCR_1&diff=123105&oldid=prevAdeline millet at 09:26, 18 September 20122012-09-18T09:26:53Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h1>PCR using a GoTaq<span class="exposant">®</span> polymerase</h1></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h1>PCR using a GoTaq<span class="exposant">®</span> polymerase</h1></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h2>Goal</h2></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h2>Goal</h2></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> Amplify a section of DNA.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> Amplify a <ins class="diffchange diffchange-inline">specific </ins>section of DNA.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></section></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></section></div></td></tr>
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</table>Adeline millethttp://2012.igem.org/wiki/index.php?title=Team:Grenoble/Biology/Protocols/PCR_1&diff=100231&oldid=prevGreghansen at 13:20, 7 September 20122012-09-07T13:20:30Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <tr><th><i><div class="petit"><center>SAFETY AND USEFUL RECOMMANDATION</center></div></i></th></tr></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <tr><th><i><div class="petit"><center>SAFETY AND USEFUL RECOMMANDATION</center></div></i></th></tr></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <tr><td><div class="petit">It was noticed that it exists a certain number of different thermocycler models.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <tr><td><div class="petit">It was noticed that it exists a certain number of different thermocycler models.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <del class="diffchange diffchange-inline">That is why biologists do not know, all the time, how the </del>device <del class="diffchange diffchange-inline">they will use, </del>has <del class="diffchange diffchange-inline">to </del>be <del class="diffchange diffchange-inline">manipulated</del>. <del class="diffchange diffchange-inline">This</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <ins class="diffchange diffchange-inline">Each </ins>device has <ins class="diffchange diffchange-inline">its own user guide which should </ins>be <ins class="diffchange diffchange-inline">thoroughly read before using the device</ins>. </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del>can present risks <del class="diffchange diffchange-inline">for thermocycler users</del>. Indeed <del class="diffchange diffchange-inline">a thermocycler </del>is an electric device which can become very hot,</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> The use of Thermocyclers </ins>can present risks. Indeed <ins class="diffchange diffchange-inline">it </ins>is an electric device which can become very hot,</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> and in case of misused, people can be injured<del class="diffchange diffchange-inline">. Consequently it is very important for users to know how to use</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> and in case of misused, people can be injured.</div></td></tr></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> the device. It is recommended to read the operating instructions before using it. Besides, the device has protective measures to prevent</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> accidents</del>.</div></td></tr></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </table></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </table></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </center></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </center></div></td></tr>
</table>Greghansenhttp://2012.igem.org/wiki/index.php?title=Team:Grenoble/Biology/Protocols/PCR_1&diff=95916&oldid=prevAdeline millet at 09:28, 5 September 20122012-09-05T09:28:36Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <tr><td><div class="petit">It was noticed that it exists a certain number of different thermocycler models.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <tr><td><div class="petit">It was noticed that it exists a certain number of different thermocycler models.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> That is why biologists do not know, all the time, how the device they will use, has to be manipulated. This</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> That is why biologists do not know, all the time, how the device they will use, has to be manipulated. This</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> can <del class="diffchange diffchange-inline">cause </del>risks for thermocycler users. Indeed a thermocycler is an electric device which can become hot,</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> can <ins class="diffchange diffchange-inline">present </ins>risks for thermocycler users. Indeed a thermocycler is an electric device which can become <ins class="diffchange diffchange-inline">very </ins>hot,</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> and in case of misused people can be injured. Consequently it is very important for users to know how to use</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> and in case of misused<ins class="diffchange diffchange-inline">, </ins>people can be injured. Consequently it is very important for users to know how to use</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> the device. It is recommended to read the operating <del class="diffchange diffchange-inline">instruction </del>before using it<del class="diffchange diffchange-inline">. In our group we managed to</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> the device. It is recommended to read the operating <ins class="diffchange diffchange-inline">instructions </ins>before using it. Besides<ins class="diffchange diffchange-inline">, </ins>the device <ins class="diffchange diffchange-inline">has protective measures </ins>to prevent</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> learn how to use it before doing any manipulation</del>. Besides <del class="diffchange diffchange-inline">on </del>the device <del class="diffchange diffchange-inline">there are also protections </del>to prevent</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <ins class="diffchange diffchange-inline">accidents</ins>.</div></td></tr></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <del class="diffchange diffchange-inline">accident</del>.</div></td></tr></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </center></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </center></div></td></tr>
</table>Adeline millethttp://2012.igem.org/wiki/index.php?title=Team:Grenoble/Biology/Protocols/PCR_1&diff=93322&oldid=prevAdeline millet at 14:50, 3 September 20122012-09-03T14:50:39Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h2>Protocol</h2></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h2>Protocol</h2></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> Following are the instructions from the</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> Following are the instructions from the</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><a href="http://www.promega.com/~/media/files/resources/protocols/product%20information%20sheets/g/gotaq%20dna%20polymerase%20m300.pdf?la=en">Promega website</a>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><a href="http://www.promega.com/~/media/files/resources/protocols/product%20information%20sheets/g/gotaq%20dna%20polymerase%20m300.pdf?la=en<ins class="diffchange diffchange-inline">" target="blank</ins>">Promega website</a>.</div></td></tr>
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</table>Adeline millethttp://2012.igem.org/wiki/index.php?title=Team:Grenoble/Biology/Protocols/PCR_1&diff=93309&oldid=prevAdeline millet at 14:47, 3 September 20122012-09-03T14:47:34Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>Set up the thermocycler at the right temperatures.</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>Set up the thermocycler at the right temperatures.</li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <br/></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <li>In a <del class="diffchange diffchange-inline">50 </del>µL PCR tube, introduce : <ul><div class="petit"></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <li>In a <ins class="diffchange diffchange-inline">200 </ins>µL PCR tube, introduce : <ul><div class="petit"></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>1.25 µL of forward primer (10µM)</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>1.25 µL of forward primer (10µM)</li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>1.25 µL of reverse primer (10µM)</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>1.25 µL of reverse primer (10µM)</li></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <li>Put the <del class="diffchange diffchange-inline">50 </del>µL PCR tube into the thermocycler.</li></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <li>Put the <ins class="diffchange diffchange-inline">200 </ins>µL PCR tube into the thermocycler.</li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <br/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>Turn on the device.</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>Turn on the device.</li></div></td></tr>
</table>Adeline millethttp://2012.igem.org/wiki/index.php?title=Team:Grenoble/Biology/Protocols/PCR_1&diff=93308&oldid=prevAdeline millet at 14:46, 3 September 20122012-09-03T14:46:20Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <tr><th><i><div class="petit"><center>SAFETY AND USEFUL RECOMMANDATION</center></div></i></th></tr></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <tr><th><i><div class="petit"><center>SAFETY AND USEFUL RECOMMANDATION</center></div></i></th></tr></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <tr><td><div class="petit">Those products are not dangerous at first sight, but in case of spreading</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <tr><td><div class="petit">Those products are not dangerous at first sight, but in case of spreading</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> or <del class="diffchange diffchange-inline">in case of products on hands or on </del>any <del class="diffchange diffchange-inline">parts on </del>the body, it is recommended to wash this part and</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> or <ins class="diffchange diffchange-inline">contacts with </ins>any <ins class="diffchange diffchange-inline">part of </ins>the body, it is recommended to wash this part and</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> to dry it after. <del class="diffchange diffchange-inline">Be careful to use </del>for each sample <del class="diffchange diffchange-inline">a different sterile cone</del>. It is also important to</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> to dry it after. <ins class="diffchange diffchange-inline">Use a different pipette tip </ins>for each sample. It is also important to</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <del class="diffchange diffchange-inline">well </del>close tubes after putting the <del class="diffchange diffchange-inline">ingredient in</del>. <del class="diffchange diffchange-inline">Moreover the </del>DNA <del class="diffchange diffchange-inline">cannot resist until </del>it is not</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> close tubes after putting the <ins class="diffchange diffchange-inline">different reagents</ins>. DNA <ins class="diffchange diffchange-inline">is more sensitive to its environment while </ins>it is not <ins class="diffchange diffchange-inline">circulared </ins>in a plasmid and <ins class="diffchange diffchange-inline">introduced</ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> incorporated </del>in a plasmid and into a cell. Therefore <del class="diffchange diffchange-inline">there is no chance that there are </del>any <del class="diffchange diffchange-inline">possible</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> </ins>into a cell. Therefore<ins class="diffchange diffchange-inline">, a spreading has very few chances to lead to </ins>any <ins class="diffchange diffchange-inline">bad </ins>consequences.</div></td></tr></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del>consequences <del class="diffchange diffchange-inline">because of a leakage in the environment</del>.</div></td></tr></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </table></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </table></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </center></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </center></div></td></tr>
</table>Adeline millethttp://2012.igem.org/wiki/index.php?title=Team:Grenoble/Biology/Protocols/PCR_1&diff=92645&oldid=prevAdeline millet at 08:55, 3 September 20122012-09-03T08:55:40Z<p></p>
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<td colspan='2' style="background-color: white; color:black;">← Older revision</td>
<td colspan='2' style="background-color: white; color:black;">Revision as of 08:55, 3 September 2012</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><section></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><section></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <h1>PCR using a GoTaq polymerase</h1></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <h1>PCR using a GoTaq<ins class="diffchange diffchange-inline"><span class="exposant">®</span> </ins>polymerase</h1></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h2>Goal</h2></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h2>Goal</h2></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> Amplify a section of DNA.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> Amplify a section of DNA.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>1.25 µL of reverse primer (10µM)</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>1.25 µL of reverse primer (10µM)</li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>2.5 µL of dNTP (2 mM)</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>2.5 µL of dNTP (2 mM)</li></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <li>5 µL of GoTaq Reaction Buffer</li></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <li>5 µL of GoTaq<ins class="diffchange diffchange-inline"><span class="exposant">®</span> </ins>Reaction Buffer</li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>variable volume of template DNA</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>variable volume of template DNA</li></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <li>0.12 µL of GoTaq polymerase</li></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <li>0.12 µL of GoTaq<ins class="diffchange diffchange-inline"><span class="exposant">®</span> </ins>polymerase</li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>to 25 µL of nuclease-free water</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>to 25 µL of nuclease-free water</li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></div></td></tr>
</table>Adeline millethttp://2012.igem.org/wiki/index.php?title=Team:Grenoble/Biology/Protocols/PCR_1&diff=92642&oldid=prevAdeline millet: Created page with "{{:Team:Grenoble/Templates/Biology}} <html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en"> <body id="Biology"> <div id="cadre"> <section> <h1>PCR using a GoTaq..."2012-09-03T08:54:32Z<p>Created page with "{{:Team:Grenoble/Templates/Biology}} <html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en"> <body id="Biology"> <div id="cadre"> <section> <h1>PCR using a GoTaq..."</p>
<p><b>New page</b></p><div>{{:Team:Grenoble/Templates/Biology}}<br />
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<h1>PCR using a GoTaq polymerase</h1><br />
<h2>Goal</h2><br />
Amplify a section of DNA.<br />
</section><br />
<br/><br />
<section><br />
<h2>Protocol</h2><br />
Following are the instructions from the<br />
<a href="http://www.promega.com/~/media/files/resources/protocols/product%20information%20sheets/g/gotaq%20dna%20polymerase%20m300.pdf?la=en">Promega website</a>.<br />
<br/><br />
<br/><br />
<ol><br />
<li>Set up the thermocycler at the right temperatures.</li><br />
<br/><br />
<li>In a 50 µL PCR tube, introduce : <ul><div class="petit"><br />
<li>1.25 µL of forward primer (10µM)</li><br />
<li>1.25 µL of reverse primer (10µM)</li><br />
<li>2.5 µL of dNTP (2 mM)</li><br />
<li>5 µL of GoTaq Reaction Buffer</li><br />
<li>variable volume of template DNA</li><br />
<li>0.12 µL of GoTaq polymerase</li><br />
<li>to 25 µL of nuclease-free water</li><br />
</div><br />
</ul><br />
<br/><br />
<center><br />
<table><br />
<tr><th><i><div class="petit"><center>SAFETY AND USEFUL RECOMMANDATION</center></div></i></th></tr><br />
<tr><td><div class="petit">Those products are not dangerous at first sight, but in case of spreading<br />
or in case of products on hands or on any parts on the body, it is recommended to wash this part and<br />
to dry it after. Be careful to use for each sample a different sterile cone. It is also important to<br />
well close tubes after putting the ingredient in. Moreover the DNA cannot resist until it is not<br />
incorporated in a plasmid and into a cell. Therefore there is no chance that there are any possible<br />
consequences because of a leakage in the environment.</div></td></tr><br />
</table><br />
</center><br />
<br/><br />
<li>Put the 50 µL PCR tube into the thermocycler.</li><br />
<br/><br />
<li>Turn on the device.</li><br />
<br/><br />
<center><br />
<table><br />
<tr><th><i><div class="petit"><center>SAFETY AND USEFUL RECOMMANDATION</center></div></i></th></tr><br />
<tr><td><div class="petit">It was noticed that it exists a certain number of different thermocycler models.<br />
That is why biologists do not know, all the time, how the device they will use, has to be manipulated. This<br />
can cause risks for thermocycler users. Indeed a thermocycler is an electric device which can become hot,<br />
and in case of misused people can be injured. Consequently it is very important for users to know how to use<br />
the device. It is recommended to read the operating instruction before using it. In our group we managed to<br />
learn how to use it before doing any manipulation. Besides on the device there are also protections to prevent<br />
accident.</div></td></tr><br />
</table><br />
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{{:Team:Grenoble/menu}}</div>Adeline millet