Team:Wellesley TheTech/Project/Wetlab

BAC PACK

• TEAM
  • Members
  • Notebooks
  • Acknowledgements
• PROJECT
  • Overview
  • Digital Component
  • Wetlab Component
  • Background
  • Hardware
• OUTREACH
  • Practices
  • Methodology
  • Collaboration
• MEDAL FULFILLMENT

WETLAB COMPONENT

Description of above image

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Goal

The goal of the wetlab component of BacPack is to connect the digital experience to a hands-on, lab-based one. In the digital activity visitors explore designing bacteria to have a variety of new, useful functions for helping humans survive in different extreme environments. In the wetlab activity visitors get try their hands at the same real-world process of engineering a bacteria to have a new property of their choosing.

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Activity Design Outline

Visitors transform a living bacteria to make it do something new:
-5 DNA options that change the physical appearance of bacteria in different ways
-Each ‘color’ represents a bacteria function that can provide one of the 5 needs on Mars
-Visitors pick the DNA they want to use and transform e.coli with it
-They can see a photo of their grown ‘mars bacteria’ several days later

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Development

Plasmids:
Selected 5 bioBricks from the iGEM distribution kit:
-Lux Operon (BBa_K325909) - Bioluminescence pathway
amilGFP (BBa_K592010) - Yellow chromoprotein
aeBlue (BBa_K864401) - Blue chromoprotein
eforRed (BBa_K592012) - Red/pink chromoprotein
asPink (BBa_K1033933) - Pink/purple chromoprotein

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Transformation

Optimized to be accessible/easy for walk-up museum visitors of all ages and skill levels:
Reagents (pre-made by staff):
 - 6cm LB-agar plates with 25 ug/ml Chloramphenicol
 - Plasmid DNA in 100ul CaCl2
 - 20 ul competent bacteria
Transformation protocol (done by visitors):
 - Combine 100 DNA with 20 ul of competent bacteria
 - 30 sec on ice
 - 40 sec heat shock at 42 degrees C
 - Plate out bacteria and put in incubator