User:Elaine Haykin/Notebook/American University Chemistry Lab/2013/01/31
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Note: Step 3 was skipped.
into the appropriate number of 14-ml BD Falcon polypropylene roundbottom tubes. Prepare an additional 100-μl aliquot of cells for use as a transformation control.
fresh 1:10 dilution of a 14.2 M stock solution of β-ME (diluted in distilled water), to each polypropylene tube containing the competent cells, for a final concentration of 25 mM β-ME. Swirl the tubes gently.
gently. For the control transformation reaction, add 1 μl of the pUC18 control plasmid to a separate 100-μl aliquot of the competent cells and swirl gently.
Preheat SOC medium (see Preparation of Media and Reagents) in a
42°C water bath for use in step 10.
Heat-pulse each transformation reaction in a 42°C water bath for
45 seconds. The duration of the heat pulse is critical for optimal transformation efficiencies.
Incubate the reactions on ice for 2 minutes.
reaction and incubate the reactions at 37°C for 1 hour with shaking at 225–250 rpm.