Publication No. 10526
Student Laboratory Kit
Materials Included In Kit
Microcentrifuge tubes, 10
Additional Materials Required
Water, distilled, 20–30 mL
Aliquot 200 μL of loading dye solution into each of the ten microcentrifuge tubes. Practice the entire pipetting procedure so that you can demonstrate the well-loading technique with skill for your students.
Wear chemical splash goggles, chemical-resistant gloves and a chemical-resistant apron. Please review current Safety Data Sheets for additional safety, handling and disposal information.
Please consult your current Flinn Scientific Catalog/Reference Manual for general guidelines and specific procedures, and review all federal, state and local regulations that may apply, before proceeding. Liquid materials from this lab can be disposed of according to Flinn Suggested Disposal Method #26b. Gels can be rinsed, dried, and stored for future use.
Correlation to Next Generation Science Standards (NGSS)†
Science & Engineering PracticesPlanning and carrying out investigations
Disciplinary Core IdeasMS-PS1.A: Structure and Properties of Matter
MS-PS2.B: Types of Interactions
MS-LS1.A: Structure and Function
HS-PS1.A: Structure and Properties of Matter
HS-PS2.B: Types of Interactions
HS-LS1.A: Structure and Function
Crosscutting ConceptsStructure and function
Electrophoresis test materials are usually expensive or available only in small quantity. The tests require the materials to be transferred with a micropipet and precise techniques. Practice these pipetting techniques with simulated materials before experimenting with real test materials.
The visualization and characterization of molecules, such as DNA, RNA and proteins, rely on a vital technique—electrophoresis. Electrophoresis is an integral step in many “hot” research areas, such as DNA fingerprinting and sequencing. Electrophoresis is a process by which charged particles (ions) move in response to an electrical force. Electrophoretic separation may be performed in free solution (moving–boundary electrophoresis) or in a solid matrix (zone electrophoresis), such as agarose or polyacrylamide gels. This exercise will focus on a practice technique commonly used in agarose gel electrophoresis.
Water, distilled, 20–30 mL
Watch carefully as your instructor demonstrates the well-loading technique using the specific micropipets available in your lab.
Wear chemical splash goggles, chemical-resistant gloves and a chemical-resistant apron. Wash hands thoroughly with soap and water before leaving the laboratory.
1. Add enough water to the top of the practice gel station so that the entire surface is submerged in water. The water simulates the buffer solution that is used in an actual electrophoresis chamber.
2. Using a micropipet, practice loading 10 μL of loading dye into one row of practice wells. The goal is to place 10 μL of loading dye into the bottom of each well without poking any holes into the practice gel. (The practice gel is actually more rigid than real agarose.) Once the loading dye has been drawn into the pipet tip, carefully introduce the tip to the end of the well pointing towards the middle of the well itself. The tip should break the surface of the water directly over the end of the well. Slowly expel the practice loading dye. Be careful not to push the tip into the bottom of the well. If the well is mistakenly punctured with real agarose, the sample will be lost. Practice loading dye contains sucrose, which is heavier than water and causes the sample to sink and stay in the bottom of the well.
3. Take turns using the micropipet and filling the wells.
4. Practice the technique until an entire row of wells has been successfully loaded. When all wells in a practice station are filled, rinse the practice station under slow running water, being careful not to splash any dye onto clothing or other objects.
5. The rinsed station can be used again for another practice round, if desired.
6. Consult your instructor for appropriate disposal procedures.