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The BioBuilder Lab Experience: Eau That Smell!

Explore the population growth curve of bacteria and compare different designs to see which generates the strongest banana smell. Learn about synthetic biology and molecular genetics techniques.

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The BioBuilder Lab Experience: Eau That Smell!

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  1. The BioBuilder Lab Experience:Eau That Smell!

  2. PRESENT • Where can it fit? • Population Ecology • The Big Idea: Compare competing designs • Objectives: • Explore how synthetic biology differs from genetic engineering. • Investigate and measure the population growth curve of bacteria. • Employ microbiology techniques. • Properly use synthetic biology and molecular genetics terms. • Microbiology • Molecular Genetics • Enrichment

  3. Design Build Test BioBuilder Emphasis:An Engineering Paradigm The focus of this lab

  4. What do we know? Parts buildDevices E. coli exhibit a predictable growth curve (lag, log, stat) The Eau d’coli device designed by the 2006 MIT iGEM team expresses the banana smell during the stationary phase.

  5. Questions to Consider 1. What happens in the original design? 2. What is the role of the promoter? 3. What is the role of the ATF1 ORF? 4. What is the role of the ATF1 enzyme?

  6. Your Challenge... We have been sent 4 strains of E. coli. Each has a different composition of parts. Test each banana generating device to see which will generate the strongest smell. smells like bananas at stationary phase smells like bananas when not in stationary smells like bananas when in log phase no banana smell

  7. 1-1 1-1 1-4 1-3 1-4 1-1 1-2 1-4 1-3 1-2 1-3 1-2 PREPARATION Sequence for Protocol A (Protocol B is an alternative where students can prepare cultures of each strain and take samples every 20 minutes... this requires a full day and many classes participating): 1. Prepare cultures of each strain (see prep videos on preparing Liquid Overnights and Stock Cultures). 2. Place a sample in the refrigerator before it has time to incubate. Growth Curve These will be the LAGphase samples. 3. Let each sample grow for 6 hours on a magnetic stir plate at room temperature. Cell Density These will be the LOGphase samples. 4. Let remaining sample run overnight on a magnetic stir plate at room temperature. Time (h) These will be the STATIONARYphase samples. There will be 12 samples all together (4 cultures each at three time points).

  8. TODAY... For LAG and LOG samples you will measure: 1. OD 600 using the Spec 20. This will tell you the population of bacteria at LAG and LOG phase. If a Spec is NOT available, you can prepare samples on the McFarland Turbidity Scale. 2. The banana smell by comparison with the banana standards (prepared from banana oil) at LAG and LOG phase. 3. Be sure to replace the cover on the cultures after smelling. Coffee in between smells??? 0 Smell Test Cell Density 1 1-1 1-1 1-1 1-4 1-4 1-4 1-3 1-3 1-3 2 1-2 1-2 1-2 3 4 LOG LAG LAG & LOG 5 6 Banana Standards PERFORM 1 OD600 How many cells? 1:10 dilutions McFarland Standards 2 Mmmmm! How sweet it is!

  9. If you are employing Protocol B, you will need to make 4 data tables like this, one for each strain TOMORROW... 1. Repeat for STATIONARY phase samples.

  10. Submit Your Data Here:

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