Team:Macquarie Australia/Protocols/Electroporation

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Revision as of 04:53, 29 August 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