DNA Structureand Analysis

Activity 4.3: Dye Electrophoresis

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Activity 4.3: Dye Electrophoresis

Research questions:– What charge do dyes have?– Are samples composed of multiple dyes?

Objectives:– Load and run dyes on an agarose gel– Determine the charge of each

electrophoresed dye– Determine the composition of each colored

sample

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Safety Reminders

Follow all laboratory safety procedures– Appropriate PPE should be worn at all times– Attention should be given when using electricity in the

presence of liquids. If there are leaks in equipment, notify teacher immediately

– Turn off power before removing lid and ensure that lid is in place before turning power on

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Skills to Master

Set up a horizontal gel electrophoresis chamber Load an agarose gel submerged in running buffer Perform agarose gel electrophoresis Program and utilize a power supply

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Running an Agarose Gel

Play video: Agarose Gel Electrophoresis

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Student Workstation Materials

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Protocol Highlights/Tips

If you are extracting dyes from candy, extract in 5% glycerol

Extract in the smallest amount of glycerol possible to ensure dyes are concentrated

If using food coloring make a 0.25% v/v solution using 5% glycerol as diluent

Ready-made loading dyes should be used at 1x concentration

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Summary

Make sure to:– Record all steps in your notebook– Ensure all lane contents are

recorded in an organized manner in your notebook

– Record the power supply settings and times in your notebook

– Place the micropipet tip just under the surface of the running buffer and deliver the dye into the well. Do not try to get the tip into the well. It could pierce the bottom of the well or break the side