http://2008.igem.org/wiki/index.php?title=Newcastle_University_Wetlab/14_August_2008&feed=atom&action=historyNewcastle University Wetlab/14 August 2008 - Revision history2024-03-28T21:53:21ZRevision history for this page on the wikiMediaWiki 1.16.5http://2008.igem.org/wiki/index.php?title=Newcastle_University_Wetlab/14_August_2008&diff=79247&oldid=prevMtaschuk at 16:27, 28 October 20082008-10-28T16:27:36Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Thursday 14th August==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Thursday 14th August==</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>* BBa_I746101 and BBa_746107 were [[isolated]] and run on a [[gel]] together with the 4 restrictions performed 13.08.08.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>* BBa_I746101 and BBa_746107 were [[<ins class="diffchange diffchange-inline">Isolating Plasmid from Cells (Miniprep)|</ins>isolated]] and run on a [[<ins class="diffchange diffchange-inline">Agarose Gel Electrophoresis|</ins>gel]] together with the 4 restrictions performed 13.08.08.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>'''Lane 1:''' 1kb ladder</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>'''Lane 1:''' 1kb ladder</div></td></tr>
</table>Mtaschukhttp://2008.igem.org/wiki/index.php?title=Newcastle_University_Wetlab/14_August_2008&diff=47241&oldid=prevMtaschuk: New page: {{:Team:Newcastle_University/WetLabCalendarHeader}} ==Thursday 14th August== * BBa_I746101 and BBa_746107 were isolated and run on a gel together with the 4 restrictions performe...2008-09-21T18:29:44Z<p>New page: {{:Team:Newcastle_University/WetLabCalendarHeader}} ==Thursday 14th August== * BBa_I746101 and BBa_746107 were <a href="/Isolated" title="Isolated">isolated</a> and run on a <a href="/Gel" title="Gel">gel</a> together with the 4 restrictions performe...</p>
<p><b>New page</b></p><div>{{:Team:Newcastle_University/WetLabCalendarHeader}}<br />
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==Thursday 14th August==<br />
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* BBa_I746101 and BBa_746107 were [[isolated]] and run on a [[gel]] together with the 4 restrictions performed 13.08.08.<br />
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'''Lane 1:''' 1kb ladder<br />
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'''Lane 2:''' empty<br />
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'''Lane 3:''' pGFPrrnB restricted with EcoRI + NheI<br />
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'''Lane 4:''' pJWV021 restricted with NheI + BglII<br />
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'''Lane 5:''' pUC57-ncl08 restricted with EcoRI + NheI<br />
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'''Lane 6:''' pUC57-ncl08 restricted with NheI + BglII<br />
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'''Lane 7:''' BBa_746101<br />
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'''Lane 8:''' BBa_746107<br />
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The gel showed that there was very little pGFPrrnB (restricted with EcoRI + NheI) and no pJWV021. It was concluded that the 2 step restriction and purification method reduced the plasmid yield significantly. As it was later discovered that this method was unnecessary (as NheI and BglII are compatible using buffer M), it was decided to perform the restriction as a single step in future.<br />
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The gel also showed successful restrictions of the pUC57-ncl08 plasmid and lots of the two BioBrick samples.<br />
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* The 2.2kb fragments from the pUC57-ncl08 restrictions were cut out of the gel using a UV light box and scalpel blade (see [[Cutting a Specific Band from Agarose Gel]]). The resulting gel slices were then purified to remove all gel and enzymes (see [[Purifying DNA from Gel Slices]]).</div>Mtaschuk