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Team:Alberta/Notebook/TransformingCells
From 2010.igem.org
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(→TSS Transformation Protocol) |
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The TSS transformation was tested. The procedure is listed here. | The TSS transformation was tested. The procedure is listed here. | ||
| - | + | <html><a href="http://www.pnas.org/content/86/7/2172.abstract">One-step preparation of competent Escherichia coli: transformation and storage of bacterial cells in the same solution</a></html> | |
| - | solution | + | |
*Acquisition of reagents- June 3, 2010 | *Acquisition of reagents- June 3, 2010 | ||
*Preparation of buffer - June 7, 2010 | *Preparation of buffer - June 7, 2010 | ||
*Cell preparation - June 7, 2010 | *Cell preparation - June 7, 2010 | ||
| - | * | + | *Protocol trial - June 10, 2010 |
| - | *Assessment of efficiency - | + | *Assessment of efficiency - June 11, 2010 |
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| + | Results: | ||
| + | Transformation with pSB1C3 gave a transformation efficiency of 4.8*10^7 colonies/&muL. | ||
==Rubidium chloride competent cells and lyophilization== | ==Rubidium chloride competent cells and lyophilization== | ||
Revision as of 02:13, 28 October 2010
Transforming Cells
Project Timeline: Click on an image to see more information
Transformation typically requires expensive equipment and storing competent cells is difficult for many high schools who do not have -80oC freezers. We tried a few methods of transforming cells and making them competent to attempt to solve this problem in our kit.
TSS Transformation Protocol
The TSS transformation was tested. The procedure is listed here.
- Acquisition of reagents- June 3, 2010
- Preparation of buffer - June 7, 2010
- Cell preparation - June 7, 2010
- Protocol trial - June 10, 2010
- Assessment of efficiency - June 11, 2010
Results: Transformation with pSB1C3 gave a transformation efficiency of 4.8*10^7 colonies/&muL.
Rubidium chloride competent cells and lyophilization
This method was adapted from a method used by Justin Fedor.
- Preparation of transformation buffer - July 6, 2010
- Preparation of cells - July 10, 2010
- Preparing competent cells - July 11, 2010
- Lyophilization - July 13, 2010
- Transformation of FD cells - July 14, 2010
- Preparation of cells 2 - July 30, 2010
- Lyophilization - August 2, 2010
- Transformation - August 5, 2010
