Team:Washington/Notebook

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Protocols

Protein Gel

Microscope

Flow Cytometry

Supernatant Protein Purification, 50mL

Ni-column Set up

Supernatant Protein Purification, 2mL

Gene Synthesis

Colony PCR

Gene Assembly using the NheI and PstI sites

Gene Assembly using the NheI site


Project Time Line

  • Winter Quarter
    • Introduction to iGEM
    • Synthetic Biology Seminar
    • Plan for project ideas
  • Spring Quarter
    • Narrow down potential projects
    • Choose Project
    • Order oligos and start synthesizing genes
    • Obtained Funding
    • Stock Lab
  • June
    • Sequence genes
    • Preliminary binding assays for biotinylated fluorophore
    • Introduction to Fold-it as a tool for protein design
    • Test target proteins for solubility and expression
    • Start assembly of secretion genes
  • July
    • Develop and perform assays for testing legacy surface display bio-bricks
    • Transfer prtDEF contig from secretion system into low copy plasmid
    • Test target proteins for functionality
  • August
    • Finish assembly of secretion system
    • Start cell lines containing various forms of secretion system, make competent
    • Transform competent cells containing secretion system with target vector
    • Test for secretion of target protein
    • Start design of new display construct
  • September
    • Switch secretion system into new cell line, make cells competent
    • Transform with target vector
    • Test for secretion
    • Start Presentation
    • Start t-shirt design
    • Insert streptavidin into new display vector
  • October
    • Fine tune secretion assay, adjust controls
    • Finalize characterization of legacy parts
    • Practice presentation
    • Characterize target bio-bricks
    • Prepare for Jamboree