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Team:Yale/Our Project/Protocols/transformation
From 2010.igem.org
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<h4> Standard Transformation Protocol </h4> | <h4> Standard Transformation Protocol </h4> | ||
| - | 1. Preheat <a id="link" href="https://2010.igem.org/Team:Yale/Our_Project/Protocols/ | + | 1. Preheat <a id="link" href="https://2010.igem.org/Team:Yale/Our_Project/Protocols/SOC_broth">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/> | ||
| Line 35: | Line 35: | ||
6. Heat pulse tubes in 37°C degree water fro 30-40 seconds<br/> | 6. Heat pulse tubes in 37°C degree water fro 30-40 seconds<br/> | ||
7. Incubate tubes on ice for 2 minutes<br/> | 7. Incubate tubes on ice for 2 minutes<br/> | ||
| - | 8. Add 500 μL of preheated <a id="link" href="https://2010.igem.org/Team:Yale/Our_Project/Protocols/ | + | 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/> | ||
| + | <br /> | ||
| + | <a id="nav" href="https://2010.igem.org/Team:Yale/Our_Project/Protocols"> >>> click here to view more protocols</a> | ||
<!------------- BUT NOT BELOW HERE -------------> | <!------------- BUT NOT BELOW HERE -------------> | ||
</p> | </p> | ||
Latest revision as of 03:03, 28 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.
>>> click here to view more protocols
