By engineering a reliable coldshock promoter, we can provide a switch to turn on gene expression in order to control cell protein expression using temperature.
By engineering a reliable coldshock promoter, we can provide a switch to turn on gene expression in order to control cell protein expression using temperature.
===Notebook===
===Notebook===
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January 1-15 Met for the first time during class time as an Instructional Focus. Discussed project ideas and iGEM in general. Gave out protocols to study
+
January 1-15---Met for the first time during class time as an Instructional Focus. Discussed project ideas and iGEM in general. Gave out protocols to study
-
January 16-31 Practiced transformations with p-GLO from Biorad. Learned about Bacterial Antibiotic resistance using Biorad Antibiotic Resistance Lab.
+
-
February 1-14 Discussed project ideas. Planned for visit to Georgia Institute of Technology, Styczynski Lab. Sold Pretzels for fund raiser.
+
----January 16-31---Practiced transformations with p-GLO from Biorad. Learned about Bacterial Antibiotic resistance using Biorad Antibiotic Resistance Lab.
-
February 15-28 Studied and took test over iGEM protocols and 3A Assembly. Finalized list of Biobrick parts of interest. Outlined initial ideas. Use K410000 to characterize cold shock heat generator.
+
-
March 1-14 March 9 Field trip to GATech.
+
----February 1-14---Discussed project ideas. Planned for visit to Georgia Institute of Technology, Styczynski Lab. Sold Pretzels for fund raiser.
+
+
----February 15-28---Studied and took test over iGEM protocols and 3A Assembly. Finalized list of Biobrick parts of interest. Outlined initial ideas. Use K410000 to characterize cold shock heat generator.
+
+
----March 1-14--- Field trip to GATech on March 9
+
• Rehydrated J23119, J13002, J04450, K410000
• Rehydrated J23119, J13002, J04450, K410000
+
• Miniprepped parts
• Miniprepped parts
+
• Ran diagnostic gels
• Ran diagnostic gels
+
• Sent parts for sequencing
• Sent parts for sequencing
+
• Calculated concentration of DNA
• Calculated concentration of DNA
+
• Met with team members from GATech iGEM. Discussed the K410000 project.
• Met with team members from GATech iGEM. Discussed the K410000 project.
+
• New idea for project; isolate HybB and make it a Biobrick. GATech team will send their frozen stocks.
• New idea for project; isolate HybB and make it a Biobrick. GATech team will send their frozen stocks.
-
March 15-28 Worked on research for the new parts. Wrote Project proposal.
+
-
April 1-24 April 1st:
+
----March 15-28---Worked on research for the new parts. Wrote Project proposal.
+
+
----April 1-24
+
+
April 1st:
+
• Transformed again
• Transformed again
+
• Made Amp plates
• Made Amp plates
+
• Cataloged Gaucher 2010 box
• Cataloged Gaucher 2010 box
+
• Plated the HypB with RFP on Amp plates
• Plated the HypB with RFP on Amp plates
+
April 4th:
April 4th:
+
• Liquid culture results were analyzed
• Liquid culture results were analyzed
+
• Took the HypB samples and mixed with Glycerol (Glyceroled Bio Bricks) and placed them in 80oC freezer
• Took the HypB samples and mixed with Glycerol (Glyceroled Bio Bricks) and placed them in 80oC freezer
+
• Streaked LB plates with HypB samples, promoter, and K410000 (bio brick)
• Streaked LB plates with HypB samples, promoter, and K410000 (bio brick)
+
• Made more LB plates
• Made more LB plates
+
• Miniprepped hypB parts and K410 from Gaucher box
• Miniprepped hypB parts and K410 from Gaucher box
+
• Calculated the DNA concentration
• Calculated the DNA concentration
+
• Calculated concentrations for sequencing
• Calculated concentrations for sequencing
+
• Wrote instructions for Cold Shock
• Wrote instructions for Cold Shock
+
April 24th:
April 24th:
+
• Nanodropped with DNA concentrations
• Nanodropped with DNA concentrations
+
• Ligation ( Protocol)
• Ligation ( Protocol)
+
• Digest of RFP, hypB, psBIC3, 410
• Digest of RFP, hypB, psBIC3, 410
-
May 1- May 14 • Transformation
+
+
----May 1- May 14
+
+
• Transformation
+
• RFP control and 410 RFP grew but only RFP has an expression
• RFP control and 410 RFP grew but only RFP has an expression
+
• Ran transformation efficiency test with Protocol
• Ran transformation efficiency test with Protocol
+
• Prep for experiment- plated 10 plates and inoculated 16 tubes
• Prep for experiment- plated 10 plates and inoculated 16 tubes
+
• Experiment for temperature variation was successful but did not prove hypothesis
• Experiment for temperature variation was successful but did not prove hypothesis
Revision as of 21:29, 6 June 2013
We are the Lambert High School iGEM team from Forsyth County, Georgia. We are honored to be the first high school iGEM team from Georgia, and this is our first year in the competition.
Tell us more about your project. Give us background. Use this as the abstract of your project. Be descriptive but concise (1-2 paragraphs)
We are Georgia's first high school iGEM team. We are from Lamber High School in Forsyth County, Georgia, just outside Atlanta. This is the first year that our school has had an iGEM program, and we are very excited to be part of this group.
Our instructor is Mrs. Janet Standeven, who teaches biology and environmental science at Lambert. We are also being sponsored by Dr. Mark Styczynski from Georgia Tech's Chemical Engineering department.
The members of our iGEM team are very enthusiastic about biotechnology and molecular engineering and are glad to be presented with the opportunity to gain valuable experience in this ever-expanding bio technical field and to have been able to work in doctorate labs at the prestigious Georgia Institute of Technology being just high school students.
Project
In 2010, Georgia Institute of Technology’s iGem team submitted the part K410000, a periplasmic heat generator made with HybB and OmpA and AOX. The following year several teams used HybB in their projects with mixed results. Our project is the continuation these projects and the characterization of HybB. To do this we will put RFP under HybB promotion. In addition to we will put OmpA AOX under three constituitive promoters. The purpose is to determine whether the strength of constituitive promoters will change the amount of heat released by generator OmpA AOX. With HybB we are hoping to get reliable results from cold shock treatment.
Future Aspirations
By engineering a reliable coldshock promoter, we can provide a switch to turn on gene expression in order to control cell protein expression using temperature.
Notebook
January 1-15---Met for the first time during class time as an Instructional Focus. Discussed project ideas and iGEM in general. Gave out protocols to study
January 16-31---Practiced transformations with p-GLO from Biorad. Learned about Bacterial Antibiotic resistance using Biorad Antibiotic Resistance Lab.
February 1-14---Discussed project ideas. Planned for visit to Georgia Institute of Technology, Styczynski Lab. Sold Pretzels for fund raiser.
February 15-28---Studied and took test over iGEM protocols and 3A Assembly. Finalized list of Biobrick parts of interest. Outlined initial ideas. Use K410000 to characterize cold shock heat generator.
March 1-14--- Field trip to GATech on March 9
• Rehydrated J23119, J13002, J04450, K410000
• Miniprepped parts
• Ran diagnostic gels
• Sent parts for sequencing
• Calculated concentration of DNA
• Met with team members from GATech iGEM. Discussed the K410000 project.
• New idea for project; isolate HybB and make it a Biobrick. GATech team will send their frozen stocks.
March 15-28---Worked on research for the new parts. Wrote Project proposal.
April 1-24
April 1st:
• Transformed again
• Made Amp plates
• Cataloged Gaucher 2010 box
• Plated the HypB with RFP on Amp plates
April 4th:
• Liquid culture results were analyzed
• Took the HypB samples and mixed with Glycerol (Glyceroled Bio Bricks) and placed them in 80oC freezer
• Streaked LB plates with HypB samples, promoter, and K410000 (bio brick)
• Made more LB plates
• Miniprepped hypB parts and K410 from Gaucher box
• Calculated the DNA concentration
• Calculated concentrations for sequencing
• Wrote instructions for Cold Shock
April 24th:
• Nanodropped with DNA concentrations
• Ligation ( Protocol)
• Digest of RFP, hypB, psBIC3, 410
May 1- May 14
• Transformation
• RFP control and 410 RFP grew but only RFP has an expression
• Ran transformation efficiency test with Protocol
• Prep for experiment- plated 10 plates and inoculated 16 tubes
• Experiment for temperature variation was successful but did not prove hypothesis
Results/Conclusions
What did you achieve over the course of your semester?
Safety
All members of the team participated in Lab safety training at our High School. In addition 5 members of the team completed Right to Know, General Lab Safety, Biohazard training and Recombinant DNA safety training offered by Georgia Institute of Technology.
Whenever the team was performing procedures, everybody wore safety goggles and gloves. We followed procedures very carefully, and we were circumspect with all procedures and cleanup.
We took a safety quiz at the beginning of the team formation.
We were supervised by Mrs. Standeven, Dr. Styczynski, and Mrs. Cochran the entire time we were in the Georgia Tech lab.
Attributions
Who worked on what?
Everyone in the Lambert iGem team is motivated and dedicated to enhance the team and our projects. Ms.Standeven, has dedicated many hours of her day to help prepare for upcoming labs.
Human Practices
In our inaugural year we focused on educating our teachers, fellow classmates and administration about the purpose of iGEM. We recruited members from the 9-12 grade which gave a large audience with whom to spread the information. Synthetic Biology as a topic was a new idea even to some of our teachers. Several members of our team volunteer at both Science Olympiad camps and STEM camps. They brought activities we had performed as team to the younger audiences in order to familiarize them with DNA.
Activities
Our favorite team snack is Pretzels.
King of Pops was a special treat in Atlanta on a work day.
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