Transforming chemically competent cells - Revision history

http://openwetware.org/index.php?title=Transforming_chemically_competent_cells&feed=atom&action=history Transforming chemically competent cells - Revision history 2013-05-18T07:41:18Z Revision history for this page on the wiki MediaWiki 1.13.2 http://openwetware.org/index.php?title=Transforming_chemically_competent_cells&diff=603080&oldid=prev Paul R Jaschke at 01:24, 17 May 2012 2012-05-17T01:24:38Z

<p></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 01:24, 17 May 2012</td> </tr> <tr><td colspan="2" class="diff-lineno">Line 10:</td> <td colspan="2" class="diff-lineno">Line 10:</td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Incubate cells for 30 seconds at <math>42^o</math>C.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Incubate cells for 30 seconds at <math>42^o</math>C.</div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#*Note: According to the [http://www.pubmedcentral.gov/articlerender.fcgi?tool=pubmed&pubmedid=2648393 original TSS paper] and qualitative experience ([[Josh Michener|JM]]), this step is completely optional and may actually reduce transformation efficiency.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#*Note: According to the [http://www.pubmedcentral.gov/articlerender.fcgi?tool=pubmed&pubmedid=2648393 original TSS paper] and qualitative experience ([[Josh Michener|JM]]), this step is completely optional and may actually reduce transformation efficiency.</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;">#* I tested this with DH5a Z1 and pUC19 and found that heat shock at 42C for 30 sec improved transformation efficiency 10-fold ([[User:Paul_R_Jaschke|Paul Jaschke]])</ins></div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Incubate cells on ice for 2 min.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Incubate cells on ice for 2 min.</div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Add 1 mL [[SOC]] ([[2XYT]] and LB are also suitable, original paper suggests LB + 20mM glucose) at room temp.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Add 1 mL [[SOC]] ([[2XYT]] and LB are also suitable, original paper suggests LB + 20mM glucose) at room temp.</div></td></tr>  </table>

Paul R Jaschke http://openwetware.org/index.php?title=Transforming_chemically_competent_cells&diff=368409&oldid=prev Vaishnavi Ananth at 09:31, 19 November 2009 2009-11-19T09:31:13Z

<p></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 09:31, 19 November 2009</td> </tr> <tr><td colspan="2" class="diff-lineno">Line 23:</td> <td colspan="2" class="diff-lineno">Line 23:</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>==BioCoder version==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==BioCoder version==</div></td></tr> <tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Following is the Transforming chemically competent cells protocol in BioCoder, a high-level programming language for expressing biology protocols. What you see here is the auto-generated text ouput of the protocol that was coded up in BioCoder (see Source code). More information about <del class="diffchange diffchange-inline">BioStream </del>can be found on my home page. Feel free to mail me your comments/ suggestions.[[User:Vaishnavi Ananth|Vaishnavi]]</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Following is the Transforming chemically competent cells protocol in BioCoder, a high-level programming language for expressing biology protocols. What you see here is the auto-generated text ouput of the protocol that was coded up in BioCoder (see Source code). More information about <ins class="diffchange diffchange-inline">BioCoder </ins>can be found on my home page. Feel free to mail me your comments/ suggestions.[[User:Vaishnavi Ananth|Vaishnavi]]</div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Text Output====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Text Output====</div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Transforming chemically competent cells protocol]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Transforming chemically competent cells protocol]]</div></td></tr>  </table>

Vaishnavi Ananth http://openwetware.org/index.php?title=Transforming_chemically_competent_cells&diff=368408&oldid=prev Vaishnavi Ananth at 09:30, 19 November 2009 2009-11-19T09:30:43Z

<p></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 09:30, 19 November 2009</td> </tr> <tr><td colspan="2" class="diff-lineno">Line 22:</td> <td colspan="2" class="diff-lineno">Line 22:</td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*heat shock of 42°C 45s increased efficiency 15-20 fold compared to no heat shock</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*heat shock of 42°C 45s increased efficiency 15-20 fold compared to no heat shock</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">BioStream </del>version==</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>==<ins class="diffchange diffchange-inline">BioCoder </ins>version==</div></td></tr> <tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Following is <del class="diffchange diffchange-inline">theTransforming </del>chemically competent cells protocol in <del class="diffchange diffchange-inline">BioStream</del>, a high-level programming language for expressing biology protocols. What you see here is the auto-generated text ouput of the protocol that was coded up in <del class="diffchange diffchange-inline">BioStream </del>(see Source code). More information about BioStream can be found on my home page. Feel free to mail me your comments/ suggestions.[[User:Vaishnavi Ananth|Vaishnavi]]</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Following is <ins class="diffchange diffchange-inline">the Transforming </ins>chemically competent cells protocol in <ins class="diffchange diffchange-inline">BioCoder</ins>, a high-level programming language for expressing biology protocols. What you see here is the auto-generated text ouput of the protocol that was coded up in <ins class="diffchange diffchange-inline">BioCoder </ins>(see Source code). More information about BioStream can be found on my home page. Feel free to mail me your comments/ suggestions.[[User:Vaishnavi Ananth|Vaishnavi]]</div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Text Output====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Text Output====</div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Transforming chemically competent cells protocol]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Transforming chemically competent cells protocol]]</div></td></tr>  </table>

Vaishnavi Ananth http://openwetware.org/index.php?title=Transforming_chemically_competent_cells&diff=361510&oldid=prev Vaishnavi Ananth at 06:57, 23 October 2009 2009-10-23T06:57:56Z

<p></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 06:57, 23 October 2009</td> </tr> <tr><td colspan="2" class="diff-lineno">Line 21:</td> <td colspan="2" class="diff-lineno">Line 21:</td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*using SOC instead of LB + 20mM glucose increased efficiency 3 fold</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*using SOC instead of LB + 20mM glucose increased efficiency 3 fold</div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*heat shock of 42°C 45s increased efficiency 15-20 fold compared to no heat shock</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*heat shock of 42°C 45s increased efficiency 15-20 fold compared to no heat shock</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;">==BioStream version==</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;">Following is theTransforming chemically competent cells protocol in BioStream, a high-level programming language for expressing biology protocols. What you see here is the auto-generated text ouput of the protocol that was coded up in BioStream (see Source code). More information about BioStream can be found on my home page. Feel free to mail me your comments/ suggestions.[[User:Vaishnavi Ananth|Vaishnavi]]</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;">====Text Output====</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;">[[Transforming chemically competent cells protocol]]</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;">====Source Code====</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;">[[Transforming chemically competent cells protocol - source code]]</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>[[Category:Protocol]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Category:Protocol]]</div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Category:Escherichia coli]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Category:Escherichia coli]]</div></td></tr>  </table>

Vaishnavi Ananth http://openwetware.org/index.php?title=Transforming_chemically_competent_cells&diff=288223&oldid=prev Torsten Waldminghaus at 17:10, 23 February 2009 2009-02-23T17:10:50Z

<p></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 17:10, 23 February 2009</td> </tr> <tr><td colspan="2" class="diff-lineno">Line 1:</td> <td colspan="2" class="diff-lineno">Line 1:</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;">{{back to protocols}}</ins></div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>''Also see [[Preparing chemically competent cells]]''</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>''Also see [[Preparing chemically competent cells]]''</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>

Torsten Waldminghaus http://openwetware.org/index.php?title=Transforming_chemically_competent_cells&diff=183805&oldid=prev Hugh Kingston at 15:47, 4 February 2008 2008-02-04T15:47:08Z

<p></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 15:47, 4 February 2008</td> </tr> <tr><td colspan="2" class="diff-lineno">Line 19:</td> <td colspan="2" class="diff-lineno">Line 19:</td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*'''[[User:Hugh Kingston|hugh kingston]] 09:46, 4 February 2008 (CST)''':I tried a few variations on this protocol, and found</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*'''[[User:Hugh Kingston|hugh kingston]] 09:46, 4 February 2008 (CST)''':I tried a few variations on this protocol, and found</div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*using SOC instead of LB + 20mM glucose increased efficiency 3 fold</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*using SOC instead of LB + 20mM glucose increased efficiency 3 fold</div></td></tr> <tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>*heat shock of 42°C 45s increased efficiency 15-20 fold</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*heat shock of 42°C 45s increased efficiency 15-20 fold <ins class="diffchange diffchange-inline">compared to no heat shock</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>[[Category:Protocol]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Category:Protocol]]</div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Category:Escherichia coli]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Category:Escherichia coli]]</div></td></tr>   </table>

Hugh Kingston http://openwetware.org/index.php?title=Transforming_chemically_competent_cells&diff=183803&oldid=prev Hugh Kingston at 15:46, 4 February 2008 2008-02-04T15:46:29Z

<p></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 15:46, 4 February 2008</td> </tr> <tr><td colspan="2" class="diff-lineno">Line 19:</td> <td colspan="2" class="diff-lineno">Line 19:</td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*'''[[User:Hugh Kingston|hugh kingston]] 09:46, 4 February 2008 (CST)''':I tried a few variations on this protocol, and found</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*'''[[User:Hugh Kingston|hugh kingston]] 09:46, 4 February 2008 (CST)''':I tried a few variations on this protocol, and found</div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*using SOC instead of LB + 20mM glucose increased efficiency 3 fold</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*using SOC instead of LB + 20mM glucose increased efficiency 3 fold</div></td></tr> <tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>*heat shock of <del class="diffchange diffchange-inline">either </del>42°C 45s increased efficiency 15-20 fold</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*heat shock of 42°C 45s increased efficiency 15-20 fold</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>[[Category:Protocol]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Category:Protocol]]</div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Category:Escherichia coli]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Category:Escherichia coli]]</div></td></tr>  </table>

Hugh Kingston http://openwetware.org/index.php?title=Transforming_chemically_competent_cells&diff=183802&oldid=prev Hugh Kingston at 15:46, 4 February 2008 2008-02-04T15:46:10Z

<p></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 15:46, 4 February 2008</td> </tr> <tr><td colspan="2" class="diff-lineno">Line 16:</td> <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>#Grow overnight at 37 &deg;C.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Grow overnight at 37 &deg;C.</div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Save the rest of the transformants in liquid culture at 4 &deg;C.  If nothing appears on your plate, you can spin this down, resuspend in enough medium to spread on one plate and plate it all. This way you will find even small numbers of transformants.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Save the rest of the transformants in liquid culture at 4 &deg;C.  If nothing appears on your plate, you can spin this down, resuspend in enough medium to spread on one plate and plate it all. This way you will find even small numbers of transformants.</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;">*'''[[User:Hugh Kingston|hugh kingston]] 09:46, 4 February 2008 (CST)''':I tried a few variations on this protocol, and found</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;">*using SOC instead of LB + 20mM glucose increased efficiency 3 fold</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;">*heat shock of either 42°C 45s increased efficiency 15-20 fold</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>[[Category:Protocol]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Category:Protocol]]</div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Category:Escherichia coli]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Category:Escherichia coli]]</div></td></tr>  </table>

Hugh Kingston http://openwetware.org/index.php?title=Transforming_chemically_competent_cells&diff=183519&oldid=prev Hugh Kingston at 17:09, 2 February 2008 2008-02-02T17:09:39Z

<p></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 17:09, 2 February 2008</td> </tr> <tr><td colspan="2" class="diff-lineno">Line 10:</td> <td colspan="2" class="diff-lineno">Line 10:</td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#*Note: According to the [http://www.pubmedcentral.gov/articlerender.fcgi?tool=pubmed&pubmedid=2648393 original TSS paper] and qualitative experience ([[Josh Michener|JM]]), this step is completely optional and may actually reduce transformation efficiency.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#*Note: According to the [http://www.pubmedcentral.gov/articlerender.fcgi?tool=pubmed&pubmedid=2648393 original TSS paper] and qualitative experience ([[Josh Michener|JM]]), this step is completely optional and may actually reduce transformation efficiency.</div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Incubate cells on ice for 2 min.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Incubate cells on ice for 2 min.</div></td></tr> <tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#Add 1 mL [[SOC]] ([[2XYT]] and LB are also suitable) at room temp.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Add 1 mL [[SOC]] ([[2XYT]] and LB are also suitable<ins class="diffchange diffchange-inline">, original paper suggests LB + 20mM glucose</ins>) at room temp.</div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Incubate for 1 hour at <math>37^o</math>C on shaker.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Incubate for 1 hour at <math>37^o</math>C on shaker.</div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#*Note: Can also save some time here by reducing incubation to ~45 min.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#*Note: Can also save some time here by reducing incubation to ~45 min.</div></td></tr>  </table>

Hugh Kingston http://openwetware.org/index.php?title=Transforming_chemically_competent_cells&diff=90438&oldid=prev Reshma P. Shetty at 22:45, 5 December 2006 2006-12-05T22:45:31Z

<p></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 22:45, 5 December 2006</td> </tr> <tr><td colspan="2" class="diff-lineno">Line 17:</td> <td colspan="2" class="diff-lineno">Line 17:</td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Save the rest of the transformants in liquid culture at 4 &deg;C.  If nothing appears on your plate, you can spin this down, resuspend in enough medium to spread on one plate and plate it all. This way you will find even small numbers of transformants.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Save the rest of the transformants in liquid culture at 4 &deg;C.  If nothing appears on your plate, you can spin this down, resuspend in enough medium to spread on one plate and plate it all. This way you will find even small numbers of transformants.</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>[[Category:<del class="diffchange diffchange-inline">E.Coli </del>Protocol]]</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>[[Category:Protocol<ins class="diffchange diffchange-inline">]]</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">[[Category:Escherichia coli</ins>]]</div></td></tr>  </table>

Reshma P. Shetty