Berglund:Chemical Transformation

From OpenWetWare

Jump to: navigation, search

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

Personal tools