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Team:Macquarie Australia/Protocols/Electroporation
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Pipette 1µL diluted (1:3) Gibson product into fresh eppendorf tube<br><br> | Pipette 1µL diluted (1:3) Gibson product into fresh eppendorf tube<br><br> | ||
Add 50µL competent cells (stir very gently to avoid shear)<br><br> | Add 50µL competent cells (stir very gently to avoid shear)<br><br> | ||
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Dry electric cuvette<br><br> | Dry electric cuvette<br><br> | ||
Pipette 50 µL competent cells to electric cuvette<br><br> | Pipette 50 µL competent cells to electric cuvette<br><br> | ||
Revision as of 06:41, 13 September 2013
Electroporation
(Keep materials on ice !)
Pipette 1µL diluted (1:3) Gibson product into fresh eppendorf tube
Add 50µL competent cells (stir very gently to avoid shear)
Dry electric cuvette
Pipette 50 µL competent cells to electric cuvette
Pulse cuvette
Settings:presets/bacteria/e-coli/1mm/1800V/25uf/200Ώ)
Quickly add 500µL LB broth (warmed in hot water bath) to cuvette
Mix by gently pipetting up and down
Working quickly, transfer contents to fresh tube
Incubate in warm room under agitation one hour
