Team:Berkeley Wetlab/Passenger: MGFP

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MGFP-5 BCA Protein Quantification Assay

This assay tests for the presence of mgfp-5 displayed on the surface of E. coli through its ability to bind to the bottom of polystyrene plates.

Constructs used:

  • mgfp-5 + spacers + displayers (79)
  • mgfp-5 + displayers (14)
  • displayers only (15)

Experiment run with 5x replicate of each condition

Day 1: Grow cells

  • Pick cells, using pin tool, with appropriate constructs from frozen stock plates into 96-well blocks with 1 ml of LB+AC media
  • Place blocks in 37C shaker overnight to grow to saturation

Day 2: Inducing

  • Set up 96-well blocks with 1 ml of LB+AC+arabinose media:
    • Add 1 ul of arabinose at a concentration of (100mg/ml) to one ml of LB+AC media (1: 1000)
  • Take plates with constructs out of the 37C incubator and add 10 ul of each construct to the LB+AC+arabinose media to induce
  • Grow


Day 3: Incubate in Artificial Seawater

Day 4: Run BCA protein quantification assay