A protocoll outlining the Preperation and transformation of electro-competent cells for Vibrio natriegensPreparation of electrocompetent cells (Vibrio natrigens)10 mL BHI (Brain haert infusion) + v2 salts Overnight cultureInoculation of a new BHI + v2 media with 1% of the overnight culture as inoculum.Grow at 37°C shaking to an OD of 0.5Chill the culture on ice for 15 minUse a chilled (4°C) centrifuge at 4,500 r.p.m. for 20 minDecant the supernatant diligently and carefullyResuspend gently in 5-10 mL Electroporation-Buffer (680 mM sucrose, 7 mM K2HPO4, pH 7), then fill the falcon tube to the top.To wash, spin again at 4.500 r.p.m. for 15 min at 4 °C, then resuspend gently in 5 mL Electroporation-Buffer. Repeat washing twice for a total of three times.Spin again at 4.500 r.p.m. for 15 min at 4 °C, decant supernatant and resuspend carefully in the residual buffer.Adjust volume for a OD of 16Aliquot in chilled Micro-Reaction-Tubes, then snap freeze in liquid nitrogenStore at –80°C Electroporation remove one aliquot with electrocompetent cells from storagekeep on ice until thawedadd plasmid and mix gentlyTransfer to chilled electroporation cuvetteElectroporation is done with the following parameters: 700(–900) V, 25 μF, 200 Ω in a 1mm cuvetteRecover for one hour at 37°C in brain heart infusion with v2 saltsPlate on LB2 agar plates. Electroporationremove one aliquot with electrocompetent cells from storagekeep on ice until thawedadd plasmid and mix gentlyTransfer to chilled electroporation cuvetteElectroporation is done with the following parameters: 700(–900) V, 25 μF, 200 Ω in a 1mm cuvetteRecover for one hour at 37°C in brain heart infusion with v2 saltsPlate on LB2 agar plates.