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Team:Macquarie Australia/Protocols/electrotransformation
From 2012.igem.org
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Dilute Assembly mix (1 µL) in water (3 µL). 1 µL was taken and mixed with 40 µL electrocompetent cells.</p> | Dilute Assembly mix (1 µL) in water (3 µL). 1 µL was taken and mixed with 40 µL electrocompetent cells.</p> | ||
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| - | <p>Defrost cells and immediately add 40 uL to 1 uL of diluted assembly mix. Sit on ice 0.5-1min. Add to 0.1cm electroporation cuvette prechilled on ice. Put in electroporator and electroporate at 1.8kV (hold both buttons down "CHG 9" appears then when you hear the beep the electroporation has occurred.) Immediately add 1 mL of SOC and mix and transfer to 15 mL tube. Grow out for 1 hr at 37 C and plate onto selective media.</p> | + | <p>Defrost cells and immediately add 40 uL to 1 uL of diluted assembly mix. Sit on ice 0.5-1min. Add to 0.1cm electroporation cuvette prechilled on ice. Put in electroporator and electroporate at 1.8kV<!(hold both buttons down "CHG 9" appears then when you hear the beep the electroporation has occurred.)!>. Immediately add 1 mL of SOC and mix and transfer to 15 mL tube. Grow out for 1 hr at 37 C and plate onto selective media.</p> |
Latest revision as of 12:10, 26 September 2012
Electrotransformation
Used Bioline electrocompetent cells.
Dilute Assembly mix (1 µL) in water (3 µL). 1 µL was taken and mixed with 40 µL electrocompetent cells.
Defrost cells and immediately add 40 uL to 1 uL of diluted assembly mix. Sit on ice 0.5-1min. Add to 0.1cm electroporation cuvette prechilled on ice. Put in electroporator and electroporate at 1.8kV. Immediately add 1 mL of SOC and mix and transfer to 15 mL tube. Grow out for 1 hr at 37 C and plate onto selective media.
