Team:Caltech/week4

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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 vector backbone from pKS001 with Biobrick prefix & suffix overhangs
  • 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 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
  • 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