http://2011.igem.org/wiki/index.php?title=Team:Cambridge/Protocols/Transformation_of_E.Coli&feed=atom&action=historyTeam:Cambridge/Protocols/Transformation of E.Coli - Revision history2024-03-29T11:44:41ZRevision history for this page on the wikiMediaWiki 1.16.0http://2011.igem.org/wiki/index.php?title=Team:Cambridge/Protocols/Transformation_of_E.Coli&diff=159301&oldid=prevHjking734 at 20:24, 21 September 20112011-09-21T20:24:11Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Transformation of E.coli==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Transformation of E.coli==</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>A simple method of transforming competent E.coli cells with your DNA of choice.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>A simple method of transforming competent E.coli cells with your DNA of choice.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><sup>[http://www.ias.ac.in/currsci/dec102002/1376.pdf]</sup> '''Mechanism of artificial transformation of E. coli with plasmid DNA – Clues from the influence of ethanol'''</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><sup>[http://www.ias.ac.in/currsci/dec102002/1376.pdf]</sup> '''Mechanism of artificial transformation of E. coli with plasmid DNA – Clues from the influence of ethanol'''</div></td></tr>
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</table>Hjking734http://2011.igem.org/wiki/index.php?title=Team:Cambridge/Protocols/Transformation_of_E.Coli&diff=149712&oldid=prevMarta kkk: /* Transformation of E.Coli */2011-09-21T07:48:22Z<p><span class="autocomment">Transformation of E.Coli</span></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>==Transformation of E.<del class="diffchange diffchange-inline">Coli</del>==</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>==Transformation of E.<ins class="diffchange diffchange-inline">coli</ins>==</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>A simple method of transforming competent E.coli cells with your DNA of choice.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>A simple method of transforming competent E.coli cells with your DNA of choice.</div></td></tr>
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</table>Marta kkkhttp://2011.igem.org/wiki/index.php?title=Team:Cambridge/Protocols/Transformation_of_E.Coli&diff=121783&oldid=prevHjking734 at 14:20, 15 September 20112011-09-15T14:20:40Z<p></p>
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<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;">__NOTOC__</del></div></td><td colspan="2"> </td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Transformation of E.Coli==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Transformation of E.Coli==</div></td></tr>
</table>Hjking734http://2011.igem.org/wiki/index.php?title=Team:Cambridge/Protocols/Transformation_of_E.Coli&diff=56738&oldid=prevJ3harvey: /* Practice */2011-07-28T09:30:36Z<p><span class="autocomment">Practice</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>:*Thaw competent E.coli cells ( 50 &mu;l in an eppendorf tube, grown to an OD<sub>600</sub> of 0.2 - 0.5) on ice. Let them sit on ice for at least 10 minutes.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>:*Thaw competent E.coli cells ( 50 &mu;l in an eppendorf tube, grown to an OD<sub>600</sub> of 0.2 - 0.5) on ice. Let them sit on ice for at least 10 minutes.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>:*Add 1 &mu;l of plasmid and mix gently by stirring with the pipette tip <del class="diffchange diffchange-inline">(</del>Use more plasmid solution if it is very weak. We have had success with about 50 ng of plasmid DNA<del class="diffchange diffchange-inline">)</del>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>:*Add 1 &mu;l of plasmid and mix gently by stirring with the pipette tip<ins class="diffchange diffchange-inline">. </ins>Use more plasmid solution if it is very weak. We have had success with about 50 ng of plasmid DNA.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>:*Incubate the cells with the DNA at 42&ordm;C for 1 minute.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>:*Incubate the cells with the DNA at 42&ordm;C for 1 minute.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>:*Add 250 &mu;l of sterile liquid broth.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>:*Add 250 &mu;l of sterile liquid broth.</div></td></tr>
</table>J3harveyhttp://2011.igem.org/wiki/index.php?title=Team:Cambridge/Protocols/Transformation_of_E.Coli&diff=56736&oldid=prevJ3harvey: /* Transformation of E.Coli */2011-07-28T09:30:05Z<p><span class="autocomment">Transformation of E.Coli</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Transformation of E.Coli==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Transformation of E.Coli==</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>A simple method of transforming competent E.coli cells with your DNA of choice.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>A simple method of transforming competent E.coli cells with your DNA of choice.</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: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">===Preparation===</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;">Often you will have to do some steps well in advance of carrying out a protocol.</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;">In this case you must take liquid broth to be autoclaved in good time. You should also turn on the water bath and incubator so they are at the required temperature when you need them, but make sure you turn them off after too.</del></div></td><td colspan="2"> </td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Theory===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Theory===</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>It is believed that prior to incubation DNA may interact with lipopolysaccharides and then cross the 'least barrier path' at the zones of adhesion described above.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>It is believed that prior to incubation DNA may interact with lipopolysaccharides and then cross the 'least barrier path' at the zones of adhesion described above.</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;">===Preparation===</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;">Ensure that you have enough sterilized liquid broth (or another medium such as SOC) and agar plates containing the correct antibiotic. Before you begin have a waterbath set to 42<sup>o</sup>C and an incubator at 37<sup>o</sup>C.</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>===Practice===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Practice===</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Take care not to disturb the competent E.coli<del class="diffchange diffchange-inline">, in other words </del>do not vortex them or <del class="diffchange diffchange-inline">place </del>them <del class="diffchange diffchange-inline">on ice</del>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Take care not to disturb the competent E.coli<ins class="diffchange diffchange-inline">: </ins>do not vortex them or <ins class="diffchange diffchange-inline">pipette </ins>them <ins class="diffchange diffchange-inline">up and down</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></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">To a clean Eppendorf tube add</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">{| border="1px"</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">|Reagent</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">|Quantity (&mu;l)</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">|-</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">|Competent E.coli cells</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">|50</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">|-</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">|DNA</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">|10</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">|-</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">|}</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">Incubate the cells with the DNA at 42&ordm;C for 1 minute.</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">Add 250&mu;l of liquid broth (which should have already been autoclaved)</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;"></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>Incubate at <del class="diffchange diffchange-inline">37</del>&ordm;<del class="diffchange diffchange-inline">c </del>for 1 <del class="diffchange diffchange-inline">hour</del>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">:*Thaw competent E.coli cells ( 50 &mu;l in an eppendorf tube, grown to an OD<sub>600</sub> of 0.2 - 0.5) on ice. Let them sit on ice for at least 10 minutes.</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 1 &mu;l of plasmid and mix gently by stirring with the pipette tip (Use more plasmid solution if it is very weak. We have had success with about 50 ng of plasmid 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">:*</ins>Incubate <ins class="diffchange diffchange-inline">the cells with the DNA </ins>at <ins class="diffchange diffchange-inline">42</ins>&ordm;<ins class="diffchange diffchange-inline">C </ins>for 1 <ins class="diffchange diffchange-inline">minute.</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 250 &mu;l of sterile liquid broth</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: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">Plate 10&mu;l on agar containing an antibiotic (the antibiotic for which resistance </del>is <del class="diffchange diffchange-inline">included in </del>the <del class="diffchange diffchange-inline">DNA being taken up by </del>the <del class="diffchange diffchange-inline">cells) and also plate 100&mu;l on another identical plate</del>. <del class="diffchange diffchange-inline">This allows for possibly overcrowding as </del>the <del class="diffchange diffchange-inline">next step after this protocol usually requires a single colony </del>to <del class="diffchange diffchange-inline">be selected</del>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">At this stage it </ins>is <ins class="diffchange diffchange-inline">important to maintain sterility to avoid contaminating </ins>the <ins class="diffchange diffchange-inline">liquid broth or </ins>the <ins class="diffchange diffchange-inline">plates</ins>. <ins class="diffchange diffchange-inline">Do not allow </ins>the <ins class="diffchange diffchange-inline">pipette tip </ins>to <ins class="diffchange diffchange-inline">touch anything before pipetting the liquid broth. Leaving the liquid broth bottle on the bench overnight should reveal any contamination in the morning</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: #ffa; color:black; font-size: smaller;"><div>Incubate overnight at 37&ordm;C</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&ordm;C for 1 hour.</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">:*Plate 10&mu;l on agar containing an antibiotic (the antibiotic for which the plasmid confers resistance) and also plate 100&mu;l on another identical plate. This allows for possible overcrowding so that single colonies can be selected.</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">:*</ins>Incubate overnight at 37&ordm;C<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>Always keep agar plates upside down so that drips of condensation and falling debris does not contaminate them.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Always keep agar plates upside down so that drips of condensation and falling debris does not contaminate them.</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>===Safety===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Safety===</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>All equipment (including gloves) that may have come into contact with the bacteria must be autoclaved <del class="diffchange diffchange-inline">for decontamination purposes</del>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>All equipment (including gloves) that may have come into contact with the bacteria must be autoclaved.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===References===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===References===</div></td></tr>
</table>J3harveyhttp://2011.igem.org/wiki/index.php?title=Team:Cambridge/Protocols/Transformation_of_E.Coli&diff=56637&oldid=prevJ3harvey: /* Theory */2011-07-28T08:44:34Z<p><span class="autocomment">Theory</span></p>
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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: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">[[#artificial_transformation|</del>Artificial transformation<del class="diffchange diffchange-inline">]] </del>is a process whereby E.coli are made competent and take up DNA from their surroundings. The main steps are chilling the cells to 0<sup>o</sup><del class="diffchange diffchange-inline">c </del>with CaCl<sub>2</sub> solution, adding the DNA and <del class="diffchange diffchange-inline">then </del>heat shocking the bacteria for a short period of time.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Artificial transformation is a process whereby E.coli are made competent and take up DNA from their surroundings. The main steps are chilling the cells to 0<sup>o</sup><ins class="diffchange diffchange-inline">C </ins>with CaCl<sub>2</sub> solution, adding the DNA and heat shocking the bacteria for a short period of time<ins class="diffchange diffchange-inline">, allowing the cells to recover at 37<sup>o</sup>C and then plate them</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>One hypothesis for artificial transformation is that the divalent cation provided by the chilled CaCl<sub>2</sub> solution which is used in creating competent cells promotes an interaction between DNA and lipopolysaccharides which are also negatively charge. Lipopolysaccharides occur in higher densities near the parts of the outer membrane of Gram negative bacteria that are in close association with the inner membrane (zones of adhesion).</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>One hypothesis for artificial transformation is that the divalent cation provided by the chilled CaCl<sub>2</sub> solution which is used in creating competent cells promotes an interaction between DNA and lipopolysaccharides which are also negatively charge. Lipopolysaccharides occur in higher densities near the parts of the outer membrane of Gram negative bacteria that are in close association with the inner membrane (zones of adhesion).</div></td></tr>
</table>J3harveyhttp://2011.igem.org/wiki/index.php?title=Team:Cambridge/Protocols/Transformation_of_E.Coli&diff=51151&oldid=prevFelix Zhou: /* Theory */2011-07-21T23:32:38Z<p><span class="autocomment">Theory</span></p>
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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: #ffa; color:black; font-size: smaller;"><div>[[#artificial_transformation|Artificial transformation]] is a process whereby E.coli are made competent and take up DNA from their surroundings. The main steps are chilling the cells to 0<sup>o</sup><del class="diffchange diffchange-inline">;</del>c with CaCl<sub>2</sub> solution, adding the DNA and then heat shocking the bacteria for a short period of time.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>[[#artificial_transformation|Artificial transformation]] is a process whereby E.coli are made competent and take up DNA from their surroundings. The main steps are chilling the cells to 0<sup>o</sup>c with CaCl<sub>2</sub> solution, adding the DNA and then heat shocking the bacteria for a short period of time.</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>One hypothesis for artificial transformation is that the divalent cation provided by the chilled CaCl<sub>2</sub> solution which is used in creating competent cells promotes an interaction between DNA and lipopolysaccharides which are also negatively charge. Lipopolysaccharides occur in higher densities near the parts of the outer membrane of Gram negative bacteria that are in close association with the inner membrane (zones of adhesion).</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>One hypothesis for artificial transformation is that the divalent cation provided by the chilled CaCl<sub>2</sub> solution which is used in creating competent cells promotes an interaction between DNA and lipopolysaccharides which are also negatively charge. Lipopolysaccharides occur in higher densities near the parts of the outer membrane of Gram negative bacteria that are in close association with the inner membrane (zones of adhesion).</div></td></tr>
</table>Felix Zhouhttp://2011.igem.org/wiki/index.php?title=Team:Cambridge/Protocols/Transformation_of_E.Coli&diff=51149&oldid=prevFelix Zhou: /* Theory */2011-07-21T23:32:26Z<p><span class="autocomment">Theory</span></p>
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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: #ffa; color:black; font-size: smaller;"><div>[[#artificial_transformation|Artificial transformation]] is a process whereby E.coli are made competent and take up DNA from their surroundings. The main steps are chilling the cells to 0 <del class="diffchange diffchange-inline">&ordm</del>; c with CaCl<sub>2</sub> solution, adding the DNA and then heat shocking the bacteria for a short period of time.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>[[#artificial_transformation|Artificial transformation]] is a process whereby E.coli are made competent and take up DNA from their surroundings. The main steps are chilling the cells to 0<ins class="diffchange diffchange-inline"><sup>o</sup></ins>;c with CaCl<sub>2</sub> solution, adding the DNA and then heat shocking the bacteria for a short period of time.</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>One hypothesis for artificial transformation is that the divalent cation provided by the chilled CaCl<sub>2</sub> solution which is used in creating competent cells promotes an interaction between DNA and lipopolysaccharides which are also negatively charge. Lipopolysaccharides occur in higher densities near the parts of the outer membrane of Gram negative bacteria that are in close association with the inner membrane (zones of adhesion).</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>One hypothesis for artificial transformation is that the divalent cation provided by the chilled CaCl<sub>2</sub> solution which is used in creating competent cells promotes an interaction between DNA and lipopolysaccharides which are also negatively charge. Lipopolysaccharides occur in higher densities near the parts of the outer membrane of Gram negative bacteria that are in close association with the inner membrane (zones of adhesion).</div></td></tr>
</table>Felix Zhouhttp://2011.igem.org/wiki/index.php?title=Team:Cambridge/Protocols/Transformation_of_E.Coli&diff=51147&oldid=prevFelix Zhou: /* Theory */2011-07-21T23:31:58Z<p><span class="autocomment">Theory</span></p>
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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: #ffa; color:black; font-size: smaller;"><div>[[#artificial_transformation|Artificial transformation]] is a process whereby E.coli are made competent and take up DNA from their surroundings. The main steps are chilling the cells to 0&ordm;c with CaCl<sub>2</sub> solution, adding the DNA and then heat shocking the bacteria for a short period of time.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>[[#artificial_transformation|Artificial transformation]] is a process whereby E.coli are made competent and take up DNA from their surroundings. The main steps are chilling the cells to 0 &ordm; c with CaCl<sub>2</sub> solution, adding the DNA and then heat shocking the bacteria for a short period of time.</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>One hypothesis for artificial transformation is that the divalent cation provided by the chilled CaCl<sub>2</sub> solution which is used in creating competent cells promotes an interaction between DNA and lipopolysaccharides which are also negatively charge. Lipopolysaccharides occur in higher densities near the parts of the outer membrane of Gram negative bacteria that are in close association with the inner membrane (zones of adhesion).</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>One hypothesis for artificial transformation is that the divalent cation provided by the chilled CaCl<sub>2</sub> solution which is used in creating competent cells promotes an interaction between DNA and lipopolysaccharides which are also negatively charge. Lipopolysaccharides occur in higher densities near the parts of the outer membrane of Gram negative bacteria that are in close association with the inner membrane (zones of adhesion).</div></td></tr>
</table>Felix Zhouhttp://2011.igem.org/wiki/index.php?title=Team:Cambridge/Protocols/Transformation_of_E.Coli&diff=51146&oldid=prevFelix Zhou: /* References */2011-07-21T23:31:32Z<p><span class="autocomment">References</span></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><sup>[http://www.ias.ac.in/currsci/dec102002/1376.pdf]</sup> Mechanism of artificial transformation of E. coli with plasmid DNA – Clues from the influence of ethanol</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><sup>[http://www.ias.ac.in/currsci/dec102002/1376.pdf]</sup> <ins class="diffchange diffchange-inline">'''</ins>Mechanism of artificial transformation of E. coli with plasmid DNA – Clues from the influence of ethanol<ins class="diffchange diffchange-inline">'''</ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{Template:Team:Cambridge/CAM_2011_TEMPLATE_FOOT}}</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{Template:Team:Cambridge/CAM_2011_TEMPLATE_FOOT}}</div></td></tr>
</table>Felix Zhou