Name __________________

MICROPIPET PRACTICE ACTIVITY

Purpose:
Most separations of proteins are on a very small scale and use micro amounts of liquids. This requires
you to manipulate a hand-held instrument called micropipette. This activity will familiarize you with
their use and gives a quick way to illustrate if you are using them properly.

Materials:
Kit with foam block holding plastic microfuge tubes of red, green, blue and yellow food dyes, 4 empty
microfuge tubes, pipette tips, filter paper, micropipette

Procedures:
A. Use of micropipette
1. Volume setting
• Look at micropipette to determine the volume range, 0.5 μl - 10 μl or 2 μl - 20 μl.
• Find out from your instructor if this pipette has a volume lock setting. If so, release the
lock as shown.
• Turn the control knob to select the needed volume. DOUBLE CHECK IT.
2. Attach a pipette tip.
3. Handle the pipette with thumb on the release button and in a vertical position. NEVER PIPET
LIQUID WITHOUT ATTACHING A TIP TO THE PIPET. NEVER LAY PIPET DOWN
WITH LIQUID IN THE TIP.
4. Filling:
• Press the control button down to the first stop.
• Immerse the pipette tip 2-mm into the liquid.
• Allow the control button to glide back slowly.
• Slide the tip out along the inside of the container.
• Wipe off any external droplets on the tip with lint-free tissue.
5. Dispensing
• Immense tip into the liquid.
• Slowly press the control button completely to activate the blow-out feature.
6. Eject the tip.

B. Practice using the pipette with samples of water to become comfortable with the feel of the
control button stops.

C. Formation of four new colors:

1. Place a 3 μl sample of the basic dye colors (red, green, blue & yellow) on filter paper and label
spots with pencil.
2. Use the chart below to mix the required mL of basic colors in correct clean, empty microtube.
3. Mix all dyes into a single droplet in the bottom of each microtube. (Use vortex mixer or clean,
plastic toothpick)
4. Place a 3 μl sample of the mixed colors on the same piece of filter paper that you placed the
basic colors.

Adapted from High School Science and Biotechnology: Conceptual Development of Electrophoresis.
<http://www.che.wsu.edu/home/modules/95modules/hardcastle/electrophoresis.html>
 Red Green Blue Yellow
Teal 5 μl 15 μl
Rose 15 μl 5 μl
Orange 6 μl 14 μl
Chartreuse 2 μl 18 μl

D. Clean up.
1. Wash the microfuge tube with the mixed colors (teal, rose, orange & chartreuse) with distilled
water.
2. Replace the clean tubes in kit for next group.
3. Clean up area.
4. Replace micropipette as indicated by your instructor.

E. Attach the filter paper this sheet of paper.

Analysis Questions:
1. Compare the sizes of dye spots on your filter paper. Are the sizes of the spots identical or
different? What does the similarity or difference tell you about your technique?

2. How do your mixed colors (teal, rose, orange and chartreuse) compare with the standard?

Adapted from High School Science and Biotechnology: Conceptual Development of Electrophoresis.
<http://www.che.wsu.edu/home/modules/95modules/hardcastle/electrophoresis.html>