"
Team:Yale/Our Project/Protocols/transformation
From 2010.igem.org
(Difference between revisions)
| Line 37: | Line 37: | ||
8. Add 500 μL of preheated <a id="link" href="https://2010.igem.org/Team:Yale/Our_Project/Protocols/SOC_broth">SOC broth</a> to each tube and incubate for an hour at 37°C with shaking<br/> | 8. Add 500 μL of preheated <a id="link" href="https://2010.igem.org/Team:Yale/Our_Project/Protocols/SOC_broth">SOC broth</a> to each tube and incubate for an hour at 37°C with shaking<br/> | ||
9. Spread 100 μL of each culture on a LB agar plate containing the appropriate antibiotic(s) and incubate overnight at 37°C <br/> | 9. Spread 100 μL of each culture on a LB agar plate containing the appropriate antibiotic(s) and incubate overnight at 37°C <br/> | ||
| + | 10. Alternatively, take 50ml tube filled with LB-media that contains the appropriate antibiotic and incubate overnight with shaking at 37°C. <br/> | ||
<!------------- BUT NOT BELOW HERE -------------> | <!------------- BUT NOT BELOW HERE -------------> | ||
</p> | </p> | ||
Revision as of 05:29, 27 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
10. Alternatively, take 50ml tube filled with LB-media that contains the appropriate antibiotic and incubate overnight with shaking at 37°C.
