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For 5 Purifications.
Packing column 45 minutes.
Restriction analysis A 35 minutes.
Restriction analysis B 50 minutes.
Gel Prep 30 minutes.
Electrophoresis 30 minutes.
Staining & Destaining 2 minutes.
In this experiment, students actually purify the restriction enzyme, Eco RI! This procedure utilizes an ion exchange chromatography step for Eco RI purification. Column fractions are assayed for the enzyme using Lambda DNA and digestion products are identified by agarose gel electrophoresis. Fractions that contain Eco RI are identified and pooled. The total and specific activities are calculated.