From 2007.igem.org

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Revision as of 15:24, 20 September 2007

Material and Reagents

 Transformation of plasmid DNA to competent E. Coli cells

        1. SOC

           2% Tryptone
           0.5% Yeast Extract
           10mM NaCl
           10mM MgSO4
           10mM MgCl2
        
        2. 1.5 mL microfuge tubes
        3. 42° C waterbath
        4. Ice
        5. 37° C shaker

Protocol

        1. Thaw competent cells on ice. 20–200µL per tube
        2. Add max. 20µL of a ligation reaction
        3. Mix very gently!
        4. Incubate the tubes on ice for 30 min
        5. Heat shock the cells for 30 sec at 42°C
        6. Place the tubes immediately on ice for at least 2 min
        7. Add 250µL of SOC medium to each tube
        8. Incubate for 1 hour at 37°C and shake vigorously
        9. Plate out the suspension on a LB agar plate containing the appropriate antibiotic.
       10. Incubate the plates overnight at 37°C