Team:Alberta/Notebook/TransformingCells

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(Difference between revisions)
(TSS Transformation Protocol)
(TSS Transformation Protocol)
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==TSS Transformation Protocol==
==TSS Transformation Protocol==
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The TSS transformation was tested. The procedure is listed here.
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The TSS transformation protocol was tested. The procedure is listed <html><a href="http://www.pnas.org/content/86/7/2172.abstract">here</a></html>.
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<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>
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*Acquisition of reagents- June 3, 2010  
*Acquisition of reagents- June 3, 2010  
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Results:
Results:
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Transformation with pSB1C3 gave a transformation efficiency of 4.8*10^7 colonies/&muL.
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Transformation with pSB1C3 gave a transformation efficiency of 4.8*10^7 colonies/μL. All colonies on the plate were strongly pink, as expected. Preparation of the cells took only 120 minutes, with a single transformation taking 90 minutes. On the other hand, a typical stock Inoue transformation cells takes a full day of work, with a single transformation taking about 120 minutes.
==Rubidium chloride competent cells and lyophilization==
==Rubidium chloride competent cells and lyophilization==

Revision as of 02:27, 28 October 2010

TEAM ALBERTA

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 protocol 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/μL. All colonies on the plate were strongly pink, as expected. Preparation of the cells took only 120 minutes, with a single transformation taking 90 minutes. On the other hand, a typical stock Inoue transformation cells takes a full day of work, with a single transformation taking about 120 minutes.

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