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Transformation Of E. Coli - With Plasmids Containing The Gene For The Green (Gfp) & Blue (Bfp) Fluorescent Proteins.

BT222

Edvotek Series 200 Experiments. For 10 transformations and controls. Time required: Transformation 45 min., Plating 5 min., Incubation overnight, Transformation efficiency 15 min.

In this experiment, transformants acquire green (gfp) or blue (bfp) fluorescent protein genes contained in two different plasmids. The gfp gene was originally isolated and cloned from the jellyfish Aequorea Victoria and was mutated to produce blue protein. In this experiment, transformed cells are selected based on antibiotic resistance. The colonies which express gfp or bfp protein are visibly green and pale blue, and will fluoresce when exposed to long wave UV light, commonly known as black light.

REQUIREMENTS: Incubation oven, 2 waterbaths, microwave oven or hot plate, Automatic pipet (5-50µl) and tips, pipet pumps or bulbs, ice, hot gloves and goggles, latex or vinyl gloves, long wave UV light, marking pens. Some components require -20ºC freezer storage.

CONTENTS: Transformation cells, Plasmid DNA for gfp and bfp, IPTG, Ampicillin Antibiotic, Calcium Chloride, ReadyPour™ Luria Broth Agar, Luria Broth Media for Recovery, Petri plates, 60 mm & 100 mm., Wrapped 10 ml pipets, Sterile Calibrated Transfer Pipets, Sterile Inoculating Loops, Microtest tubes with attached caps, Toothpicks, Complete instructions, background information, and study questions.

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