Team:UC Chile2/General Protocols

From 2012.igem.org

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<li>Directly incubate the reaction at 50°C for 1 hour</li>
<li>Directly incubate the reaction at 50°C for 1 hour</li>
<li>Transform competent cells with 8uL of assembled DNA (that leaves 4uL to check in an agarose gel if the reaction fails)</li>
<li>Transform competent cells with 8uL of assembled DNA (that leaves 4uL to check in an agarose gel if the reaction fails)</li>
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</ul>
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<h2Checking assembly</h2>
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<ul>
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<li>Check transformant colonies by colony PCR using primers for whole insert</li>
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<li>Grow positive colonies in media with corresponding antibiotic</li>
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<li>Miniprep colonies and digest plasmid with EcoRI & PstI restriction enzymes</li>
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<li>If checking protocols validate until know, proceed by sequencing plasmid</li>
</ul>
</ul>

Revision as of 04:31, 13 September 2012

Project: Luxilla - Pontificia Universidad Católica de Chile, iGEM 2012