https://bridgeslab.sph.umich.edu/protocols/index.php?action=history&feed=atom&title=PCR_Amplification_with_Platinum_Taq_High_Fidelity 2026-05-09T20:47:35Z Revision history for this page on the wiki MediaWiki 1.45.1 https://bridgeslab.sph.umich.edu/protocols/index.php?title=PCR_Amplification_with_Platinum_Taq_High_Fidelity&diff=935&oldid=prev 2015-07-17T13:43:41Z

<p>copied from surveyor assay protocol</p> <p><b>New page</b></p><div>__NOTOC__<br /> [[ Category: Molecular Biology ]]<br /> [[ Category: Cloning ]]<br /> [[ Category: Genotyping ]]<br /> <br /> ==Materials==<br /> * Platinum PCR SuperMix High Fidelity (Life Technologies cat# 12532-016)<br /> * Amplification Primers (2 uM stock solution with both primers combined): <br /> <br /> ==Amplification of Target Region==<br /> * Using Platinum Taq High Fidelity amplify region of interest (design primers, should be &lt;1000bp):<br /> * Per reaction:<br /> ** 21.5 uL Platinum Taq High Fidelity Master Mix<br /> ** 2.5 uL of Primers from a 2 uM stock solution<br /> ** 1 uL of the Extracted DNA from the previous step.<br /> * The PCR program should be<br /> ** 2 min at 95C<br /> ** 94C for 15<br /> ** 55C for 15s (adjust based on Tm of primers, use 5C less)<br /> ** 68C for 1min/kb amplicon<br /> ** Repeat steps 2-5 35X<br /> ** 68C for 10min</div>

Davebrid