BioBuilding: Synthetic Biology for Students: Lab 1: Difference between revisions
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* Culture bacteria using proper microbiology methods. | * Culture bacteria using proper microbiology methods. | ||
* Measure the growth of a bacterial population. | * Measure the growth of a bacterial population. | ||
* Define and properly use synthetic biology terms: [[http://youtu.be/X7FUTCvmiE8 | * Define and properly use synthetic biology terms: [[http://youtu.be/X7FUTCvmiE8 Part,]][[http://youtu.be/aPqqS8vIAKA Device,]] [[http://youtu.be/ER2wz3I-2KU Inverter.]] | ||
* Define and properly use molecular genetics terms: [[http://youtu.be/8oHSRBZGMdM | * Define and properly use molecular genetics terms: [[http://youtu.be/8oHSRBZGMdM Promoter,]][[http://youtu.be/8oHSRBZGMdM ribosome binding site ("RBS"),]] [[http://youtu.be/2x8pkL6bAeQ open reading frame ("ORF"),]] [[http://youtu.be/8oHSRBZGMdM Terminator,]] [[http://openwetware.org/wiki/BioBuilding:_Synthetic_Biology_for_Students:_Glossary Plasmid.]] | ||
==Introduction== | ==Introduction== | ||
[[Image:Intro lab1 BioPrimer1.png|thumb|600 px| [Media:Bioprimer no1 02.02.jpg BioPrimer #1 pdf] ]] | [[Image:Intro lab1 BioPrimer1.png|thumb|600 px| [[Media:Bioprimer no1 02.02.jpg| BioPrimer #1 pdf]] ]] | ||
For the 2006 [[http://youtu.be/5HXrrN7mV0A | For the 2006 [[http://youtu.be/5HXrrN7mV0A iGEM]] competition, MIT students designed Eau d’coli, ''E. coli'' that smell like bananas when their population is in the stationary phase. They did this by inserting a device that contains a stationary phase sensitive promoter coupled to a banana smell device, a device that contains a ribosome binding site (RBS), an open reading frame (ORF) that codes for the ATF1 enzyme and terminator sequences. The ATF1 enzyme converts isoamyl alcohol to isoamyl acetate, the molecule that gives bananas their characteristic smell. | ||
It has been suggested that a device | It has been suggested that a device to generate the banana smell during the bacteria’s log (or exponential) phase of population growth will be helpful. There are two ways to accomplish this. Both approaches will continue to use the original banana smell device but alter the function of the promoter. One approach couples the banana smell device to a new part, a log phase promoter. The other approach uses the same promoter but adds a genetic inverter. Synthetic biologists have constructed these devices for us and transformed bacteria with them.<br style="clear:both" /><br> | ||
[[Image:BioBuilding GrowthCurve Lab1.png|500px]] | [[Image:BioBuilding GrowthCurve Lab1.png|500px]] | ||
<br style="clear:both" /><br> | <br style="clear:both" /><br> | ||
We have been sent four strains of ''E. coli.'' Each contains a different device:<br> | We have been sent four strains of ''E. coli.'' Each contains a different device:<br> | ||
'''Sample 1-1.''' The original Eau | '''Sample 1-1.''' The original Eau d’coli device<br> | ||
'''Sample 1-2.''' The original Eau | '''Sample 1-2.''' The original Eau d’coli device but with an inverter added between the promoter and the RBS.<br> | ||
'''Sample 1-3.''' The banana smell generator coupled to the log phase promoter<br> | '''Sample 1-3.''' The banana smell generator coupled to the log phase promoter<br> | ||
'''Sample 1-4.''' A strain of E. coli that has no smell generating devices.<br> | '''Sample 1-4.''' A strain of ''E. coli'' that has no smell generating devices.<br> | ||
Our task will be to grow these bacterial populations and test for the banana smell as the population moves through the log phase and into the stationary phase. We will determine the population growth by using a spectrophotometer or the McFarland Turbidity Standards to measure the density of the bacteria in liquid culture. As the population increases we can assess the increasing banana smell, comparing the smell to dilutions of banana extract. | Our task will be to grow these bacterial populations and test for the banana smell as the population moves through the log phase and into the stationary phase. We will determine the population growth by using a spectrophotometer or the McFarland Turbidity Standards to measure the density of the bacteria in liquid culture. As the population increases we can assess the increasing banana smell, comparing the smell to dilutions of banana extract. | ||
==Procedure== | ==Procedure== | ||
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[[BioBuilding:_Synthetic_Biology_for_Students:_Lab_1--Protocol_B |Protocol B]] | [[BioBuilding:_Synthetic_Biology_for_Students:_Lab_1--Protocol_B |Protocol B]] | ||
When you've finished your experiments (Protocol A or Protocol B), upload your data to the BioBuilder site that's [http://www.biobuilder-submitdata.org/users/login here.] You'll be able to compare what you've measured to what other BioBuilders around the country have seen. | |||
==Lab Report== | ==Lab Report== | ||
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*[[BioBuilding: Synthetic Biology for Students: Design Assignment]] | *[[BioBuilding: Synthetic Biology for Students: Design Assignment]] | ||
*[[BioBuilding: Synthetic Biology for Students: Glossary]] | *[[BioBuilding: Synthetic Biology for Students: Glossary]] | ||
*[[http://biobuilder.org/ | *[[http://biobuilder.org/ back to biobuilder]] | ||
}} | }} |
Latest revision as of 13:29, 9 January 2012
Eau That Smell Lab |
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LAB 1: Eau that smell
Acknowledgments: This lab was developed with materials and guidance from the MIT 2006 iGEM team, as well as technical insights and help from Ginkgo BioworksObjectivesBy the conclusion of this laboratory investigation, the student will be able to:
IntroductionFor the 2006 [iGEM] competition, MIT students designed Eau d’coli, E. coli that smell like bananas when their population is in the stationary phase. They did this by inserting a device that contains a stationary phase sensitive promoter coupled to a banana smell device, a device that contains a ribosome binding site (RBS), an open reading frame (ORF) that codes for the ATF1 enzyme and terminator sequences. The ATF1 enzyme converts isoamyl alcohol to isoamyl acetate, the molecule that gives bananas their characteristic smell. It has been suggested that a device to generate the banana smell during the bacteria’s log (or exponential) phase of population growth will be helpful. There are two ways to accomplish this. Both approaches will continue to use the original banana smell device but alter the function of the promoter. One approach couples the banana smell device to a new part, a log phase promoter. The other approach uses the same promoter but adds a genetic inverter. Synthetic biologists have constructed these devices for us and transformed bacteria with them. ProcedureThere are two versions of this lab. Your teacher will inform you of the one you will be doing. Click on the links below to go to the detailed procedure: When you've finished your experiments (Protocol A or Protocol B), upload your data to the BioBuilder site that's here. You'll be able to compare what you've measured to what other BioBuilders around the country have seen. Lab ReportAs you write, be sure to define and properly use all highlighted terms throughout the introduction and other parts of the lab. I. Introduction
II. Methods
III. Results
IV. Discussion
V. Citations and references
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