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Edvotek 302 Purification of the Restriction Enzyme Eco RI
Edvotek 302 Purification of the Restriction Enzyme Eco RI

Purification of the Restriction Enzyme Eco RI - Edvotek 302

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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. Recommended for advanced courses.

For 5 purifications.

Time required:

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

Kit includes: Instructions, E. coli RY (Eco Extract (lyophilized), DEAE-Cellulose, 10x equilibration buffer, 50% glycerol, KCl, Eco RI reaction buffer, qualified water, Lambda DNA, Lambda/Eco RI marker, Eco RI dilution buffer, UltraSpec-Agarose™ powder, concentrated electrophoresis buffer, 10x gel loading solution, chromatography columns, SYBR® Safe stain, 5ml snap-top microcentrifuge tubes, 1.5 mL snap-top microcentrifuge tubes, & 50mL conical tube.

You need: Horizontal gel electrophoresis apparatus, power supply, automatic micropipette with tips, balance, water bath, ring stand and clamps, 13 x 100 mm glass test tubes, assorted laboratory glassware, permanent markers and tape, 1.5 mL microtest tubes, microwave, hot plate or burner, pipette pump, 250 mL flasks or beakers, hot gloves, safety goggles and disposable laboratory gloves, distilled or deionized water, LED transilluminator.