https://bridgeslab.sph.umich.edu/protocols/index.php?action=history&feed=atom&title=Colony_PCR 2026-05-30T23:20:57Z Revision history for this page on the wiki MediaWiki 1.45.1 https://bridgeslab.sph.umich.edu/protocols/index.php?title=Colony_PCR&diff=405&oldid=prev 2009-10-21T15:43:39Z

<p></p> <table style="background-color: #fff; color: #202122;" data-mw="interface"> <col class="diff-marker" /> <col class="diff-content" /> <col class="diff-marker" /> <col class="diff-content" /> <tr class="diff-title" lang="en"> <td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">← Older revision</td> <td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">Revision as of 15:43, 21 October 2009</td> </tr><tr><td colspan="2" class="diff-lineno" id="mw-diff-left-l28">Line 28:</td> <td colspan="2" class="diff-lineno">Line 28:</td></tr> <tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>#4C hold forever</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>#4C hold forever</div></td></tr> <tr><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>*Check PCR by Agarose Gel Electrophoresis</div></td><td class="diff-marker"></td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>*Check PCR by Agarose Gel Electrophoresis</div></td></tr> <tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;"></ins></div></td></tr> <tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">[[Category:Cloning]]</ins></div></td></tr> <tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">[[Category:PCR]]</ins></div></td></tr> <tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">[[Category:Genotyping]]</ins></div></td></tr> <tr><td colspan="2" class="diff-side-deleted"></td><td class="diff-marker" data-marker="+"></td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">[[Category:Yeast]]</ins></div></td></tr> <!-- diff cache key bridgeslabproto-mw_:diff:1.41:old-403:rev-405:php=table --> </table>

Davebrid https://bridgeslab.sph.umich.edu/protocols/index.php?title=Colony_PCR&diff=403&oldid=prev 2009-10-21T15:42:28Z

<p>Created page with &#039;==Reagents== *Primers *dNTPs (2 mM of each dNTP) *Expand polymerase (Roche) *Primers *Yeast Colonies ==Protocol== #Pick about 20% of a colony and mix in 20 uL of sterile water. ...&#039;</p> <p><b>New page</b></p><div>==Reagents==<br /> *Primers<br /> *dNTPs (2 mM of each dNTP)<br /> *Expand polymerase (Roche)<br /> *Primers<br /> *Yeast Colonies<br /> <br /> ==Protocol==<br /> #Pick about 20% of a colony and mix in 20 uL of sterile water.<br /> #Prepare PCR:<br /> *5 uL Expand Polymerase Buffer<br /> *5 uL dNTPs<br /> *1 uL of a 5X diluted stock<br /> *1 uL of a 5X diluted stock<br /> *5 uL cell suspension<br /> *0.5 uL Expand Polymerase<br /> ===Run PCR Program Colony PCR:===<br /> #94C for 5 min<br /> #94C for 10s<br /> #40C for 30s<br /> #68C for 2 min<br /> #Repeat Steps 2-4 10 times<br /> #94C for 10s<br /> #40C for 30s<br /> #68C for 1 min + 20s/cycle<br /> #Repeat Steps 6-8 19 times<br /> #68C for 7min<br /> #4C hold forever<br /> *Check PCR by Agarose Gel Electrophoresis</div>

Davebrid