In this experiment, students purify the restriction enzyme, Eco RI. This procedure utilised an ion exchange chromatograph step for Eco RI purification. Column fractions are assayed for the enzyme using Lambda DNA and digestion products are identified by gel electrophoresis. Fractions that contain Eco RI are identified and pooled. The total and specific activities are calculated. Recommended for advanced courses.
- For 5 lab groups
- Complete in 3 hours
Also required: Electrophoresis tank, power supply, UV visualisation system, water bath, microcentrifuge, UV spectrophotometer and cuvettes, automatic micropipette with tips, ring stands and clamps, and 10 ml pipettes.
Delivery information: Kit includes instructions, ion exchange matrix, chromatography columns, E.coli cell extract, equilibrium and elution buffer, Lambda DNA, Lambda/Eco RI marker, KCl, glycerol, dilution and reaction buffers, gel loading solution, agarose, electrophoresis buffer, and stain.