Team:Yale/Our Project/Protocols/transformation

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<h4> Standard Transformation Protocol </h4>
<h4> Standard Transformation Protocol </h4>
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   # Preheat SOC broth at 37 °C
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   1.  Preheat <a id="link" href="https://2010.igem.org/Team:Yale/Our_Project/Protocols/SOC_broth">SOC broth</a> at 37 °C <br/>
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   # Thaw competent cells on ice
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   2. Thaw competent cells on ice <br/>
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   # Aliquot 50-100 μL of cells into two pre-chilled 15 mL culture tubes (one for the control, one for the actual transformation)
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   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/>
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   # Add 1-2 μL of plasmid and swirl gently
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   4. Add 1-2 μL of plasmid and swirl gently <br/>
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   # Incubate tubes on ice for 30 minutes
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   5. Incubate tubes on ice for 30 minutes<br/>
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   # Heat pulse tubes in 37°C degree water fro 30-40 seconds
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   6. Heat pulse tubes in 37°C degree water fro 30-40 seconds<br/>
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   # Incubate tubes on ice for 2 minutes
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   7. Incubate tubes on ice for 2 minutes<br/>
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   # Add 500 μL of preheated SOC broth to each tube and incubate for an hour at 37°C with shaking
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   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/>
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   # Spread 100 μL of each culture on a LB agar plate containing the appropriate antibiotic(s) and incubate overnight at 37°C  
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   9. Spread 100 μL of each culture on a LB agar plate containing the appropriate antibiotic(s) and incubate overnight at 37°C <br/>
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  10. Alternatively, take 50ml tube filled with LB-media that contains the appropriate antibiotic and incubate overnight with shaking at 37°C. <br/>
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<a id="nav" href="https://2010.igem.org/Team:Yale/Our_Project/Protocols"> >>> click here to view more protocols</a>
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Latest revision as of 03:03, 28 October 2010

iGEM Yale

protocols

Standard Transformation Protocol

1. Preheat SOC broth at 37 °C
2. 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

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