Optimization the electroporation parameters of Klebsiella pneumoniae

The electroporation parameters of Klebsiella pneumoniae TUAC01 were optimized based on the electroporation efficiency.The effect of operate parameters on the electroporation efficiency was investigated,those operated parameters included pulser voltage,cell growth stage,final electro-competent cell density,EDTA level in medium and plasmid concentration.It was shown that,adding EDTA to a final concentration of 0.7mmol/L in medium,collect cell at the stage of cell density reached OD600 0.7,concentrated the electro-competent cell to a final density of larger than OD600 30 and pulser voltage selected 2kV used 2mm electroporate chamber.Under this condition,the electroporation efficiency reached to 8.58×105cfu/μg DNA transformants.