Protocols

Protein Gel

Microscopy

Flow Cytometry

Supernatant Protein Purification, 50mL

Ni-column Set up

Supernatant Protein Purification, 2mL

Gene Synthesis

Colony PCR

BioBrick Assembly using the NheI and PstI sites

BioBrick Assembly using the NheI site

SOEing PCR

Traditional Protein Purification (IMAC)

Generating a Standard curve for GFP concentration

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