DNA Structure and Analysis Activity 4.3: Dye Electrophoresis.

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Presentation transcript:

DNA Structure and Analysis Activity 4.3: Dye Electrophoresis

Biotechnology: A Laboratory Skills Course | explorer.bio-rad.com 2 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

Biotechnology: A Laboratory Skills Course | explorer.bio-rad.com 3 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

Biotechnology: A Laboratory Skills Course | explorer.bio-rad.com 4 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

Biotechnology: A Laboratory Skills Course | explorer.bio-rad.com 5 Running an Agarose Gel  Play video: Agarose Gel Electrophoresis

Biotechnology: A Laboratory Skills Course | explorer.bio-rad.com 6 Student Workstation Materials

Biotechnology: A Laboratory Skills Course | explorer.bio-rad.com 7 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

Biotechnology: A Laboratory Skills Course | explorer.bio-rad.com 8 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