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Team:Paris Saclay/Protocols/BioBrick Assembly - Revision history
2024-03-28T20:52:34Z
Revision history for this page on the wiki
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MelaCriq: /* Enzymes */
2014-10-17T21:57:02Z
<p><span class="autocomment">Enzymes</span></p>
<table style="background-color: white; color:black;">
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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 21:57, 17 October 2014</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Part A:</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Part A:</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>*Enzyme A: <del class="diffchange diffchange-inline">XbaI</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*Enzyme A: <ins class="diffchange diffchange-inline">EcoR1</ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>*Enzyme B: <del class="diffchange diffchange-inline">PstI</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*Enzyme B: <ins class="diffchange diffchange-inline">XbaI</ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Part B with '''Format II'''</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Part B with '''Format II'''</div></td></tr>
</table>
MelaCriq
http://2014.igem.org/wiki/index.php?title=Team:Paris_Saclay/Protocols/BioBrick_Assembly&diff=358531&oldid=prev
MelaCriq: /* Enzymes */
2014-10-17T21:53:18Z
<p><span class="autocomment">Enzymes</span></p>
<table style="background-color: white; color:black;">
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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 21:53, 17 October 2014</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Part B with '''Format I'''</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Part B with '''Format I'''</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>* Enzyme A: <del class="diffchange diffchange-inline">XbaI</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>* Enzyme A: <ins class="diffchange diffchange-inline">EcoRI</ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>* Enzyme B: <del class="diffchange diffchange-inline">PstI</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>* Enzyme B: <ins class="diffchange diffchange-inline">XbaI</ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>'''Part B placed before Part A:'''</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>'''Part B placed before Part A:'''</div></td></tr>
</table>
MelaCriq
http://2014.igem.org/wiki/index.php?title=Team:Paris_Saclay/Protocols/BioBrick_Assembly&diff=108757&oldid=prev
HoangVu: /* Enzymes */
2014-09-08T15:15:55Z
<p><span class="autocomment">Enzymes</span></p>
<table style="background-color: white; color:black;">
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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 15:15, 8 September 2014</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>''<u>Note:</u> Manipulate the enzymes with a proper subzero temperature support.''</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>''<u>Note:</u> Manipulate the enzymes with a proper subzero temperature support.''</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Part B placed after Part A:</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">'''</ins>Part B placed after Part A:<ins class="diffchange diffchange-inline">'''</ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Part A:</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Part A:</div></td></tr>
<tr><td colspan="2" class="diff-lineno">Line 20:</td>
<td colspan="2" class="diff-lineno">Line 20:</td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* Enzyme B: PstI</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* Enzyme B: PstI</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Part B placed before Part A:</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">'''</ins>Part B placed before Part A:<ins class="diffchange diffchange-inline">'''</ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Part A:</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Part A:</div></td></tr>
</table>
HoangVu
http://2014.igem.org/wiki/index.php?title=Team:Paris_Saclay/Protocols/BioBrick_Assembly&diff=108756&oldid=prev
HoangVu: /* Enzymes */
2014-09-08T15:15:29Z
<p><span class="autocomment">Enzymes</span></p>
<table style="background-color: white; color:black;">
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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 15:15, 8 September 2014</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Enzymes====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Enzymes====</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>''<u>Note:</u> Manipulate the enzymes with a proper subzero temperature support.''</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>''<u>Note:</u> Manipulate the enzymes with a proper subzero temperature support.''</div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">Part B placed after Part A:</ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Part A:</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Part A:</div></td></tr>
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<td colspan="2" class="diff-lineno">Line 16:</td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* Enzyme B: SpeI</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* Enzyme B: SpeI</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Part B with '''Format I''' <del class="diffchange diffchange-inline">(Part B placed after Part A):</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Part B with '''Format I'''</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>* Enzyme A<del class="diffchange diffchange-inline">: EcoRI</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>* Enzyme A: XbaI</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">* Enzyme B</del>: XbaI</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> </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">Part B with '''Format II''' (Part B placed before Part A):</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">* Enzyme A: SpeI</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: #eee; color:black; font-size: smaller;"><div>* Enzyme B: PstI</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* Enzyme B: PstI</div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">Part B placed before Part A:</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">Part A:</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">*Enzyme A: XbaI</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">*Enzyme B: PstI</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">Part B with '''Format II'''</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">* Enzyme A: EcoRI</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">* Enzyme B: SpeI</ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Digest reaction====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Digest reaction====</div></td></tr>
</table>
HoangVu
http://2014.igem.org/wiki/index.php?title=Team:Paris_Saclay/Protocols/BioBrick_Assembly&diff=60168&oldid=prev
Maschi: /* Procedure */
2014-08-06T17:32:13Z
<p><span class="autocomment">Procedure</span></p>
<table style="background-color: white; color:black;">
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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 17:32, 6 August 2014</td>
</tr><tr><td colspan="2" class="diff-lineno">Line 7:</td>
<td colspan="2" class="diff-lineno">Line 7:</td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Procedure===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Procedure===</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"></del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Enzymes====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Enzymes====</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>''<u>Note:</u> Manipulate the enzymes with a proper subzero temperature support.''</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>''<u>Note:</u> Manipulate the enzymes with a proper subzero temperature support.''</div></td></tr>
<tr><td colspan="2" class="diff-lineno">Line 25:</td>
<td colspan="2" class="diff-lineno">Line 24:</td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Digest reaction====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Digest reaction====</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Take a 1.5ml microcentrifuge tube (Eppendorf) and put:</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Take a 1.5ml microcentrifuge tube (Eppendorf) and put:</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>## <del class="diffchange diffchange-inline">xμl </del>of H<sub>2</sub>0 milliQ (complement)</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>## <ins class="diffchange diffchange-inline">x μl </ins>of H<sub>2</sub>0 milliQ (complement)</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>## <del class="diffchange diffchange-inline">5μl </del>of buffer (Fast Digest Buffer 10x)</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>## <ins class="diffchange diffchange-inline">y μl </ins>of buffer (<ins class="diffchange diffchange-inline">''</ins>Fast Digest Buffer 10x<ins class="diffchange diffchange-inline">'') (1x final concentration</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">xμl </del>of '''Part A''' DNA</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>## <ins class="diffchange diffchange-inline">z μl </ins>of '''Part A''' DNA</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>## <del class="diffchange diffchange-inline">1μl </del>of Enzyme A</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>## <ins class="diffchange diffchange-inline">1 μl </ins>of Enzyme A</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>## <del class="diffchange diffchange-inline">1μl </del>of Enzyme B</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>## <ins class="diffchange diffchange-inline">1 μl </ins>of Enzyme B</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Repeat '''step 1''' with '''Part B'''</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Repeat '''step 1''' with '''Part B'''</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Mix gently both tubes</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Mix gently both tubes</div></td></tr>
<tr><td colspan="2" class="diff-lineno">Line 36:</td>
<td colspan="2" class="diff-lineno">Line 35:</td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Segregate process====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Segregate process====</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div># <del class="diffchange diffchange-inline">part A</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div># <ins class="diffchange diffchange-inline">If the buffer used has no Loading Dye, put it with 1x final concentration.</ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Follow the [https://2014.igem.org/Team:Paris_Saclay/Protocols/Gel_Electrophoresis Electrophoresis Protocol] with the following parameters:</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Follow the [https://2014.igem.org/Team:Paris_Saclay/Protocols/Gel_Electrophoresis Electrophoresis Protocol] with the following parameters:</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>## Make a gel <del class="diffchange diffchange-inline">'''</del>0.8%<del class="diffchange diffchange-inline">''' </del>agarose</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>## Make a gel 0.8% agarose</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>## Use <del class="diffchange diffchange-inline">xxx </del>of BET concentration</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>## Use <ins class="diffchange diffchange-inline">2% </ins>of BET concentration</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>## Use a box with a long height and a double case for the DNA.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>## Use a box with a long height and a double case for the DNA.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>## Run the gel at 100V for one hour</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>## Run the gel at 100V for one hour</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div># <del class="diffchange diffchange-inline">minimal exposure </del>of UV <del class="diffchange diffchange-inline">light</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div># <ins class="diffchange diffchange-inline">Determine the weight </ins>of <ins class="diffchange diffchange-inline">the (Eppendorf) tube prior to use.</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">mass of </del>the <del class="diffchange diffchange-inline">ependorf (before / after)</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"># Minimize </ins>UV <ins class="diffchange diffchange-inline">exposure time to avoid damaging the DNA.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div># <ins class="diffchange diffchange-inline">Put </ins>the <ins class="diffchange diffchange-inline">DNA fragment with a minimal agarose gel into the tube.</ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>''<u>Note:</u> Minimize UV exposure time to avoid damaging the DNA.''</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>''<u>Note:</u> Minimize UV exposure time to avoid damaging the DNA.''</div></td></tr>
<tr><td colspan="2" class="diff-lineno">Line 49:</td>
<td colspan="2" class="diff-lineno">Line 49:</td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====DNA extration from agarose gels====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====DNA extration from agarose gels====</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Excise DNA fragment / solubilize gel slice</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Excise DNA fragment / solubilize gel slice</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">## Determine the weight of the (Eppendorf) tube prior to use.</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">## Put the DNA fragment with a minimal agarose gel into the tube.</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>## Determine the weight of the gel slice by the difference of the tube's weight.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>## Determine the weight of the gel slice by the difference of the tube's weight.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>## For each 100 mg of agarose gel < 2% add 200 μl Buffer NTI.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>## For each 100 mg of agarose gel < 2% add 200 μl Buffer NTI.</div></td></tr>
</table>
Maschi
http://2014.igem.org/wiki/index.php?title=Team:Paris_Saclay/Protocols/BioBrick_Assembly&diff=58275&oldid=prev
Maschi at 15:39, 5 August 2014
2014-08-05T15:39:00Z
<p></p>
<table style="background-color: white; color:black;">
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<col class='diff-marker' />
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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 15:39, 5 August 2014</td>
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<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">{{Team:Paris_Saclay/protocols_header}}</ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>=BioBrick Assembly=</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>=BioBrick Assembly=</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Ligation====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Ligation====</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">[https://2014.igem.org/</del>Team:Paris_Saclay/<del class="diffchange diffchange-inline">Protocols Back to the Protocols]</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">{{</ins>Team:Paris_Saclay/<ins class="diffchange diffchange-inline">protocols_footer}}</ins></div></td></tr>
</table>
Maschi
http://2014.igem.org/wiki/index.php?title=Team:Paris_Saclay/Protocols/BioBrick_Assembly&diff=57839&oldid=prev
Maschi: /* BioBrick Assembly */
2014-08-05T12:50:07Z
<p><span class="autocomment">BioBrick Assembly</span></p>
<table style="background-color: white; color:black;">
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<col class='diff-marker' />
<col class='diff-content' />
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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 12:50, 5 August 2014</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Ligation====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Ligation====</div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">[https://2014.igem.org/Team:Paris_Saclay/Protocols Back to the Protocols]</ins></div></td></tr>
</table>
Maschi
http://2014.igem.org/wiki/index.php?title=Team:Paris_Saclay/Protocols/BioBrick_Assembly&diff=57556&oldid=prev
Maschi: /* BioBrick Assembly */
2014-08-05T08:16:09Z
<p><span class="autocomment">BioBrick Assembly</span></p>
<table style="background-color: white; color:black;">
<col class='diff-marker' />
<col class='diff-content' />
<col class='diff-marker' />
<col class='diff-content' />
<tr valign='top'>
<td colspan='2' style="background-color: white; color:black;">← Older revision</td>
<td colspan='2' style="background-color: white; color:black;">Revision as of 08:16, 5 August 2014</td>
</tr><tr><td colspan="2" class="diff-lineno">Line 39:</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>## Make a gel '''0.8%''' agarose</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>## Make a gel '''0.8%''' agarose</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>## Use xxx of BET concentration</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>## Use xxx of BET concentration</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>## Use a box with a <del class="diffchange diffchange-inline">such </del>long height and a double case for the DNA.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>## Use a box with a long height and a double case for the DNA.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>## Run the gel at 100V for one hour</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>## Run the gel at 100V for one hour</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># minimal exposure of UV light</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># minimal exposure of UV light</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># mass of the ependorf (before / after)</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># mass of the ependorf (before / after)</div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">''<u>Note:</u> Minimize UV exposure time to avoid damaging the DNA.''</ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====DNA extration from agarose gels====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====DNA extration from agarose gels====</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"># Excise DNA fragment / solubilize gel slice</ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">## Determine the weight of the (Eppendorf) tube prior to use.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">## Put the DNA fragment with a minimal agarose gel into the tube.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">## Determine the weight of the gel slice by the difference of the tube's weight.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">## For each 100 mg of agarose gel < 2% add 200 μl Buffer NTI.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">## Incubate sample for 5-10 min at 50°C.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">## Vortex the sample briefly every 2-3 min until the gel slice is completely dissolved.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"># Bind DNA</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">## Place a NucleSpin Gel and PCR Clean-up Column into a Collection Tube and load up to 700 μl sample.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">## Centrifuge for 30 seconds at 11,000 x g. Discard flow-through. </ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">## Load remaining sample if necessary and repeat the centrifugation step.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"># Wash silica membrane</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">## Add 700 μl Buffer NT3 to the column. Centrifuge for 30 s at 11,000 x g. Discard flow-through.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"># Dry silica membrane</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">## Centrifuge for 1 min at 11,000 x g to remove Buffer NT3 completely.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"># Elute DNA</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">## Place the column into a new 1.5 ml micro centrifuge tube. Add 15-30 μl Buffer NE and incubate at room temperature (18-25 °C) for 1 min.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">## Centrifuge for 1 min at 11,000 x g.</ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Ligation====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Ligation====</div></td></tr>
</table>
Maschi
http://2014.igem.org/wiki/index.php?title=Team:Paris_Saclay/Protocols/BioBrick_Assembly&diff=56943&oldid=prev
Maschi: /* Procedure */
2014-08-04T16:15:14Z
<p><span class="autocomment">Procedure</span></p>
<table style="background-color: white; color:black;">
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<td colspan='2' style="background-color: white; color:black;">Revision as of 16:15, 4 August 2014</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Segregate process====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Segregate process====</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># part A</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># part A</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div># <del class="diffchange diffchange-inline">big box, </del>gel 0.8% agarose <del class="diffchange diffchange-inline">(and </del>BET concentration<del class="diffchange diffchange-inline">???????)</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div># <ins class="diffchange diffchange-inline">Follow the [https://2014.igem.org/Team:Paris_Saclay/Protocols/Gel_Electrophoresis Electrophoresis Protocol] with the following parameters:</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">1 </del>hour</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">## Make a </ins>gel <ins class="diffchange diffchange-inline">'''</ins>0.8%<ins class="diffchange diffchange-inline">''' </ins>agarose</div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">## Use xxx of </ins>BET concentration</div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#<ins class="diffchange diffchange-inline"># Use a box with a such long height and a double case for the DNA.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">## Run the gel at 100V for one </ins>hour</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># minimal exposure of UV light</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># minimal exposure of UV light</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># mass of the ependorf (before / after)</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># mass of the ependorf (before / after)</div></td></tr>
</table>
Maschi
http://2014.igem.org/wiki/index.php?title=Team:Paris_Saclay/Protocols/BioBrick_Assembly&diff=55108&oldid=prev
Maschi: /* Digest reaction */
2014-08-01T12:56:24Z
<p><span class="autocomment">Digest reaction</span></p>
<table style="background-color: white; color:black;">
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<col class='diff-marker' />
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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 12:56, 1 August 2014</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Repeat '''step 1''' with '''Part B'''</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Repeat '''step 1''' with '''Part B'''</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Mix gently both tubes</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Mix gently both tubes</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div># </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div># <ins class="diffchange diffchange-inline">Incubate at 37°C for one hour</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">- reaction</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> </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">- procedure</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> </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">- results?</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> </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># Store '''Part B''' at -20°C.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div># Store '''Part B''' at -20°C.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
</table>
Maschi