BIOTECHNOLOGY

BIOTECNOLOGÍA


2.2. RESTRICTION ENZYMES AND CLONING

INTRODUCTION TO RESTRICTION ENZYMES

This experiment’s aim is to develop the knowledge of restriction enzymes and agarose gel electrophoresis. This kit includes all material you need to conduct practice 4 times with different groups or classes. It includes introduction to the technical, practice material and teaching guide for the teacher.

The digested phage lambda DNA is supplied.

> Kit includes:
Samples, TAE, agarose and staining method.
> You need:
Horizontal electrophoresis system, power supply, micropipet and tips, balance, microwave or hot plate, white light visualization system.

icons-ref-blueER1            4 practices

DIGESTION OF PHAGE LAMBDA WITH RESTRICTION ENZYMES

This experiment’s aim is to develop the knowledge of bacteriophage lambda and agarose gel electrophoresis. This kit includes all material you need to conduct practice 4 times with different groups or classes. It includes introduction to the technical, practice material and teaching guide for the teacher.

The phage lambda DNA and enzymes for digestion are provided. These enzymes are special and allow digestions in just 10 minutes with what the practice can be done in a short time.

> Kit includes:
Samples, TAE, agarose and staining method.
> You need:
Horizontal electrophoresis system, power supply, micropipette and tips, balance, microwave or hot plate, white light visualization system.

icons-ref-blueER2            4 practices

MAPPING OF RESTRICTION SITES ON PLASMID DNA

This experiment’s aim is to develop the DNA mapping knowledge determining cleavage sites of the restriction enzymes into a plasmid.

Plasmid DNA is cut by various combinations of restriction enzymes and resolved by agarose gel electrophoresis.

> Kit includes:
Restriction enzymes, DNA plasmid samples, agarose powder, molecular weight marker, electrophoresis buffer, staining method, calibrated pipette, and microtipped transfer pipettes. 
> You need:
Horizontal electrophoresis system, power supply, micropipette and tips, balance, microwave or hot plate, white light visualization system.

icons-ref-blueER3            4 practices

PURIFICACTION OF THE RESTRICTION ENZYME Eco RI

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.

> Kit includes:
Ion exchange matrix, chromatography columns, E. coli cell extract, equilibration and elution buffer, Lambda DNA, Lambda/Eco RI Marker, KCl, glycerol, dilution and reaction buffers, agarose, electrophoresis buffer and staining method.
> You need:
Horizontal gel electrophoresis apparatus, power supply, UV visualization system, waterbath, microcentrifuge, microwave or hot plate, UV spectrophotometer and cuvettes, micropipette with tips, ring stands & clamps, 10 ml pipettes, lab glassware, ice and ice buckets.

icons-ref-blueER4            4 practices
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