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From 2012.igem.org

Biobricks

Methods:

Transformation of E. coli competent cells Experimental procedure:

• 1) Obtain competent E coli cells from -80 C.
• 2) Hold the tubes of competent cells in your fingers until they are just defrosted, and then put them on ice immediately. 100 µl is sufficient for 2 transformations.
• 3) Add 1-10 µl of ligation mix to each tube. Incubate on ice for 5 min.
• 4) Put the tubes in the 42°C bath for 30 sec and put back on ice for 2 min.
• 5) Add 200 µl of SOC media to each tube, and incubate in the 37oC shaker for 10 min or up to 1hr. I do 10 minutes if it is just a plasmid or 1 hour for ligation mixes.
• 6) For each tube of cells, spread 5 µl onto one LB (Ampicillin) plate and 50 µl onto a second LB (Ampicillin) plate, using aseptic technique. This should give you a total of four plates.
• 7) Place your plates upside-down in the 37°C incubator.