Team:Lambert GA
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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=== | ||
- | January 1-15 | + | 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 March 9 Field trip to GATech. | |
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• Rehydrated J23119, J13002, J04450, K410000 | • Rehydrated J23119, J13002, J04450, K410000 | ||
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• Miniprepped parts | • Miniprepped parts | ||
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• Ran diagnostic gels | • Ran diagnostic gels | ||
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• Sent parts for sequencing | • Sent parts for sequencing | ||
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• Calculated concentration of DNA | • Calculated concentration of DNA | ||
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• Met with team members from GATech iGEM. Discussed the K410000 project. | • Met with team members from GATech iGEM. Discussed the K410000 project. | ||
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• 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: | |
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- | April 1st: | + | |
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• Transformed again | • Transformed again | ||
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• Made Amp plates | • Made Amp plates | ||
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• Cataloged Gaucher 2010 box | • Cataloged Gaucher 2010 box | ||
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• Plated the HypB with RFP on Amp plates | • Plated the HypB with RFP on Amp plates | ||
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April 4th: | April 4th: | ||
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• Liquid culture results were analyzed | • Liquid culture results were analyzed | ||
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• 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 | ||
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• Streaked LB plates with HypB samples, promoter, and K410000 (bio brick) | • Streaked LB plates with HypB samples, promoter, and K410000 (bio brick) | ||
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• Made more LB plates | • Made more LB plates | ||
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• Miniprepped hypB parts and K410 from Gaucher box | • Miniprepped hypB parts and K410 from Gaucher box | ||
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• Calculated the DNA concentration | • Calculated the DNA concentration | ||
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• Calculated concentrations for sequencing | • Calculated concentrations for sequencing | ||
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• Wrote instructions for Cold Shock | • Wrote instructions for Cold Shock | ||
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April 24th: | April 24th: | ||
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• Nanodropped with DNA concentrations | • Nanodropped with DNA concentrations | ||
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• Ligation ( Protocol) | • Ligation ( Protocol) | ||
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• Digest of RFP, hypB, psBIC3, 410 | • Digest of RFP, hypB, psBIC3, 410 | ||
- | + | May 1- May 14 • Transformation | |
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- | • Transformation | + | |
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• RFP control and 410 RFP grew but only RFP has an expression | • RFP control and 410 RFP grew but only RFP has an expression | ||
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• Ran transformation efficiency test with Protocol | • Ran transformation efficiency test with Protocol | ||
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• Prep for experiment- plated 10 plates and inoculated 16 tubes | • Prep for experiment- plated 10 plates and inoculated 16 tubes | ||
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• 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 00:25, 7 June 2013