https://bridgeslab.sph.umich.edu/protocols/index.php?action=history&feed=atom&title=TSC-floxed_Genotyping 2026-05-10T01:04:34Z Revision history for this page on the wiki MediaWiki 1.45.1 https://bridgeslab.sph.umich.edu/protocols/index.php?title=TSC-floxed_Genotyping&diff=737&oldid=prev 2012-12-30T19:11:16Z

<p>added categories</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 19:11, 30 December 2012</td> </tr><tr><td colspan="2" class="diff-lineno" id="mw-diff-left-l12">Line 12:</td> <td colspan="2" class="diff-lineno">Line 12:</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>PCR genotyping was performed by simultaneous amplification of both wild-type Tsc1 and the deleted allele using the following three primers in a 35 cycle PCR reaction using Perkin Elmer AmpliTaq Gold: F4536, 5′-AGGAGGCCTCTTCTGCTACC-3′; R4830, 5′-CAGCTCCGACCATGAAGTG-3′; and R6548, 5′-TGGGTCCTGACCTATCTCCTA-3′ (Fig. 1D). Products were 295 bp (wild-type) and 368 bp (mutant), and were analyzed on agarose gels.</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>PCR genotyping was performed by simultaneous amplification of both wild-type Tsc1 and the deleted allele using the following three primers in a 35 cycle PCR reaction using Perkin Elmer AmpliTaq Gold: F4536, 5′-AGGAGGCCTCTTCTGCTACC-3′; R4830, 5′-CAGCTCCGACCATGAAGTG-3′; and R6548, 5′-TGGGTCCTGACCTATCTCCTA-3′ (Fig. 1D). Products were 295 bp (wild-type) and 368 bp (mutant), and were analyzed on agarose gels.</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>&lt;/blockquote&gt;</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>&lt;/blockquote&gt;</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: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:Mouse Work]]</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:DNA]]</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:Molecular Biology]]</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:mTORC1 ]]</ins></div></td></tr> <!-- diff cache key bridgeslabproto-mw_:diff:1.41:old-736:rev-737:php=table --> </table>

Davebrid https://bridgeslab.sph.umich.edu/protocols/index.php?title=TSC-floxed_Genotyping&diff=736&oldid=prev 2012-12-30T19:10:27Z

<p>Added primers and reference for TSC1 primers</p> <p><b>New page</b></p><div>==Primers==<br /> * Cre (F) GCATT ACCGG GCAAC GAGTG ATGAG<br /> * Cre (R) GAGTG AACGA ACCTG GTCGA AATCA GTGCG<br /> <br /> * TSC1 (F) AGG AGG CCT CTT CTG CTA CC<br /> * TSC1 (R) CAG CTC CGA CCA TGA AGT G<br /> <br /> ==Reference==<br /> From Kwiatkowski &#039;&#039;et al.&#039;&#039;, Hum. Mol. Genet. 11(5):525-34. 2002. A mouse model of TSC1 reveals sex-dependent lethality from liver hemangiomas, and up-regulation of p70S6 kinase activity in Tsc1 null cells. http://www.ncbi.nlm.nih.gov/pubmed/11875047:<br /> <br /> &lt;blockquote&gt;<br /> PCR genotyping was performed by simultaneous amplification of both wild-type Tsc1 and the deleted allele using the following three primers in a 35 cycle PCR reaction using Perkin Elmer AmpliTaq Gold: F4536, 5′-AGGAGGCCTCTTCTGCTACC-3′; R4830, 5′-CAGCTCCGACCATGAAGTG-3′; and R6548, 5′-TGGGTCCTGACCTATCTCCTA-3′ (Fig. 1D). Products were 295 bp (wild-type) and 368 bp (mutant), and were analyzed on agarose gels.<br /> &lt;/blockquote&gt;</div>

Davebrid