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 college level courses.
Kit includes: instructions, ion exchange matrix, chromatography columns, E. coli cell extract, equilibration & elution buffer, Lambda DNA, Lambda/Eco RI Marker, KCl, glycerol, dilution & reaction buffers, gel loading solution, agarose, electrophoresis buffer, InstaStain® Ethidium Bromide.
You need: horizontal gel electrophoresis apparatus, power supply, UV visualization system, waterbath, microcentrifuge, microwave or hot plate, UV spectrophotometer & cuvettes, automatic micropipet with tips, ring stands & clamps, 10 ml pipets, lab glassware, ice and ice buckets.
Storage: Some Components Require Freezer Storage
Packing Column 45 min. Restriction Analysis A 35 min. Restriction Analysis B 50 min. Gel Prep 30 min. Electrophoresis 30 min. Staining & Destaining 2 min.