"
Team:Alberta/Notebook/TransformingCells
From 2010.igem.org
(→TSS Transformation Protocol) |
(→Rubidium chloride competent cells and lyophilization) |
||
| Line 41: | Line 41: | ||
==Rubidium chloride competent cells and lyophilization== | ==Rubidium chloride competent cells and lyophilization== | ||
| - | This method was adapted from a | + | This method was adapted from a competent cell protocol found <html><a href="http://www.unc.edu/depts/marzluff/protocols/rubidium_competent_cells.pdf" >here</a></html>, which was suggested to us by former iGEM team member Justin Fedor. |
*Preparation of transformation buffer - July 6, 2010 | *Preparation of transformation buffer - July 6, 2010 | ||
Revision as of 02:45, 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 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 competent cell protocol found here, which was suggested to us by former iGEM team member 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
