Berglund:Chemical Transformation

Materials
5XKCM 1.25ml of 4M KCL (fc=0.5M KCl) 1.5ml of 1M CaCl2 (fc=0.15M CaCl2) 2.5ml of 1M MgCl2 (fc=0.25M MgCl2) 4.75 ml nanopure water

Competent cells of your choice For plasmid preps use: Novablue, TG1, Top10 For protein expression use: Rosetta (DE3), BL21 (DE3), BL21 (DE3) pLyseS, BL21 STAR

Method
Transformation 1µl-2µl of ligated plasmid 59µl of water 40µl of 5x KCM 100µl of competent cells Berglund:Making_Competent_Cells:_Various_Methods

incubate on ice for 20 min heat shock for 90 sec at 42°C add 80µl SOC plate all of it on LB-amp