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Team:Caltech/week4
From 2014.igem.org
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| - | <ul><li>PCR extracted backbone from pKS001 | + | <ul><li>PCR extracted vector backbone from pKS001 with Biobrick prefix & suffix overhangs</li> |
<li>PCR purification of PCR product and then 1 hour incubation at 37°C with DPN1 enzyme to remove any remaining sticky ends</li> | <li>PCR purification of PCR product and then 1 hour incubation at 37°C with DPN1 enzyme to remove any remaining sticky ends</li> | ||
<li>PCR purification of DPN1-digested product</li> | <li>PCR purification of DPN1-digested product</li> | ||
<li>Ran a gel that confirmed presence of ~3000bp-long DNA fragment in both DPN1-digested and undigested PCR products</li> | <li>Ran a gel that confirmed presence of ~3000bp-long DNA fragment in both DPN1-digested and undigested PCR products</li> | ||
| - | <li>Gibson Assembly of the PCR fragments with the geneblocks ordered 2 weeks ago to form exportDomain-comX-mNeonGreen constructs in the vector backbone</li> | + | <li>Gibson Assembly of the PCR fragments with the geneblocks ordered 2 weeks ago to form the 4 exportDomain-comX-mNeonGreen constructs (incorporating GeneIII, OmpA, PelB, and TorA as the export domains in the N-terminus) and the 1 comX-mNeonGreen-HlyA construct, all in the vector backbone</li> |
<li>Transformation of assemblies into JM109. Plated on ampicillin plates and incubated at 37°C overnight</li> | <li>Transformation of assemblies into JM109. Plated on ampicillin plates and incubated at 37°C overnight</li> | ||
</ul> | </ul> | ||
Revision as of 01:15, 8 July 2014
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