"
Team:Yale/Our Project/Protocols/transformation
From 2010.igem.org
(Difference between revisions)
m |
|||
| Line 28: | Line 28: | ||
<!------------- WRITE HERE-------------> | <!------------- WRITE HERE-------------> | ||
<h4> Standard Transformation Protocol </h4> | <h4> Standard Transformation Protocol </h4> | ||
| - | 1. Preheat SOC broth at 37 °C <br/> | + | 1. Preheat <a id="link" href="https://2010.igem.org/Team:Yale/Our_Project/Protocols/SOC">SOC broth</a> at 37 °C <br/> |
2. Thaw competent cells on ice <br/> | 2. Thaw competent cells on ice <br/> | ||
3. Aliquot 50-100 μL of cells into two pre-chilled 15 mL culture tubes (one for the control, one for the actual transformation)<br/> | 3. Aliquot 50-100 μL of cells into two pre-chilled 15 mL culture tubes (one for the control, one for the actual transformation)<br/> | ||
Revision as of 19:05, 25 October 2010
our project
protocols
Standard Transformation Protocol
1. Preheat SOC broth at 37 °C2. Thaw competent cells on ice
3. Aliquot 50-100 μL of cells into two pre-chilled 15 mL culture tubes (one for the control, one for the actual transformation)
4. Add 1-2 μL of plasmid and swirl gently
5. Incubate tubes on ice for 30 minutes
6. Heat pulse tubes in 37°C degree water fro 30-40 seconds
7. Incubate tubes on ice for 2 minutes
8. Add 500 μL of preheated SOC broth to each tube and incubate for an hour at 37°C with shaking
9. Spread 100 μL of each culture on a LB agar plate containing the appropriate antibiotic(s) and incubate overnight at 37°C
