E. Coli Chemical Transformation
Calcium Competent Transformation



 
 

  • 100 µl Comp Cells

    •    50 µl TCM
       up to 15-20 µl DNA
       On Ice 30’    		 TCM =
    10 mM Tris 7.4
    10 mM CaCl2
    10 mM MgCl2

     
    • Heat Shock 90 sec at 42 û°C (60 - 90 sec.) No shaking.
    • Add 1 ml LB, Shake at 37 û°C for 30-60 min.

      •                                                 (30 min Carb.)
                                                      (30 min Kan.)
                                                      (30-45 Chlor.)
    • Plate
    e.g.
     
      100 ml 500 µl
    .25 = 2 0.5 ml of 2 M Tris 5 µl MgCl2
    .5 1.0 ml CaCl2 5 µl CaCl2
    .5 1.0 ml MgCl2 2.5 µl Tris
    48.75 97.5 ml H2O 487.5 µl dH2O