Team:OLS Canmore Canada/Experiment

EXPERIMENT

The Summary

In recent years, the issue of plastic pollution has become an overwhelming global crisis. Only 5% of all plastics are recycled and the rest ends up in landfills or oceans. When looking for a solution to this problem, the Design Thinking methodology learned at the Berkeley Program was applied. In the engagement with recycling stakeholders, the OLS SynBio team discovered that the issue is not the recycling of plastic, but instead the inefficient sorting of these plastics.


To further understand this issue, the team participated in many community outreach events. OLS Synbio consulted with Simon Robbins, the corporate manager of a local recycling plant, who provided guidance and insight of how the recycling cycle works. OLS SynBio also met with Peter Duck, the zero waste manager for the town of Canmore. Lastly, the team went to the Alberta Recycling Conference to learn more about how plastics are recycled in the community, and how big the team’s contribution would be. Stakeholder feedback helped to pivot and refine the project.

Some members of our team at the Canmore Sorting Facility.

The Subtitle

  1. Preparation of competent B. Subtilis SCK6 cells for immediate transformation:
  2. 2 days before transformation streak out a plate (1uL/mL final Erythromycin) of SCK 6 and grow at 37℃ overnight.
  3. Prepare seed culture: inoculate a single colony from the plate into 5 mL of LB-Erythromycin (1uL/mL) in a 50 mL Falcon tube. Incubate overnight at 37℃ at 200 rpm.
  4. Take the absorbency of the seed cultures using the spec. The OD600 should be approximately 1.0.
  5. Place 1000µL of overnight culture into a sterile Eppendorf tube for each transformation. Centrifuge and pour off the supernatant. Repeat 4 more times, to centrifuge the entire 5 mL of the overnight culture into 1 Eppendorf tube - pellet should be large. Do not discard the overnight falcon tube.
  6. Place 5 mL of Fresh LB broth (pre-warmed to 37℃) into the original overnight Falcon tube. Add 5 µL of erythromycin. Add 250 µL of 10% sterile xylose solution to the LB broth. Add 500 µL of this broth mixture to centrifuged cells in the Eppendorf tube (pipette up and down to resuspend cells). Transfer 500 µL of cells back into large Falcon tube. Incubate at 37℃ for 2 hours on the rotary shaking table.