iGEM 2009 — Team Washington

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# Team:Washington/Notebook

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* [Home](/Team%3AWashington "Team:Washington")
* [Project Description](/Team%3AWashington/Project "Team:Washington/Project")
  + [Target](/Team%3AWashington/Project/Target "Team:Washington/Project/Target")
  + [Secretion](/Team%3AWashington/Project/Secretion "Team:Washington/Project/Secretion")
  + [Display](/Team%3AWashington/Project/Display "Team:Washington/Project/Display")
* [Future Directions](/Team%3AWashington/Future "Team:Washington/Future")
  + [CDS](/Team%3AWashington/Project/CDS "Team:Washington/Project/CDS")
  + [Foldit](/Team%3AWashington/Project/FoldIt "Team:Washington/Project/FoldIt")
* **Notebook**
* [Safety](/Team%3AWashington/Safety "Team:Washington/Safety")
* [References](/Team%3AWashington/Bibliography "Team:Washington/Bibliography")
* [The Team](/Team%3AWashington/Team "Team:Washington/Team")
* [Accomplishments & Submitted BioBricks](/Team%3AWashington/Accomplishments "Team:Washington/Accomplishments")

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# **Protocols**

[Protein Gel](/Team%3AWashington/Notebook/protein_gel "Team:Washington/Notebook/protein gel")

[Microscopy](/Team%3AWashington/Notebook/Microscope "Team:Washington/Notebook/Microscope")

[Flow Cytometry](/Team%3AWashington/Notebook/Flow_Cytometry "Team:Washington/Notebook/Flow Cytometry")

[Supernatant Protein Purification, 50mL](/Team%3AWashington/Notebook/50mL_purification "Team:Washington/Notebook/50mL purification")

[Ni-column Set up](/Team%3AWashington/Notebook/Ni-column "Team:Washington/Notebook/Ni-column")

[Supernatant Protein Purification, 2mL](/Team%3AWashington/Notebook/2mL_purification "Team:Washington/Notebook/2mL purification")

[Gene Synthesis](/Team%3AWashington/Notebook/gene_synthesis "Team:Washington/Notebook/gene synthesis")

[Colony PCR](/Team%3AWashington/Notebook/colony_PCR "Team:Washington/Notebook/colony PCR")

[BioBrick Assembly using the NheI and PstI sites](/Team%3AWashington/Notebook/NheI_PstI "Team:Washington/Notebook/NheI PstI")

[BioBrick Assembly using the NheI site](/Team%3AWashington/Notebook/NheI "Team:Washington/Notebook/NheI")

[SOEing PCR](/Team%3AWashington/Notebook/SOEingPCR "Team:Washington/Notebook/SOEingPCR")

[Traditional Protein Purification (IMAC)](/Team%3AWashington/Notebook/IMAC_protocol "Team:Washington/Notebook/IMAC protocol")

[Generating a Standard curve for GFP concentration](/Team%3AWashington/Notebook/Standard_curve "Team:Washington/Notebook/Standard curve")

# **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

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