Team:Caltech/week4

From 2014.igem.org

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<ul><li>PCR extracted backbone from pKS001 plasmid with overhangs for Biobrick prefixes & suffixes</li>
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    <li>PCR purification of PCR product and then 1 hour incubation at 37&deg;C with DPN1 enzyme to remove any remaining sticky ends</li>
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    <li>PCR purification of DPN1-digested product</li>
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    <li>Ran a gel that confirmed presence of ~3000bp-long DNA fragment in both DPN1-digested and undigested PCR products</li>
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    <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>
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    <li>Transformation of assemblies into JM109. Plated on ampicillin plates and incubated at 37&deg;C overnight</li>
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Revision as of 01:13, 8 July 2014


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Notebook

Overview

Week 1

Week 2

Week 3

Week 4

Week 5

Week 6

Week 7

Week 8

Week 9

Week 10

Week 11

Week 12

Week Four

Monday, 7/7/14

  • PCR extracted backbone from pKS001 plasmid with overhangs for Biobrick prefixes & suffixes
  • PCR purification of PCR product and then 1 hour incubation at 37°C with DPN1 enzyme to remove any remaining sticky ends
  • PCR purification of DPN1-digested product
  • Ran a gel that confirmed presence of ~3000bp-long DNA fragment in both DPN1-digested and undigested PCR products
  • Gibson Assembly of the PCR fragments with the geneblocks ordered 2 weeks ago to form exportDomain-comX-mNeonGreen constructs in the vector backbone
  • Transformation of assemblies into JM109. Plated on ampicillin plates and incubated at 37°C overnight

Tuesday, 7/8/14

Wednesday, 7/9/14


Thursday, 7/10/14

Friday, 7/11/14