You sank my ... bacteriophage?
Abstract: Practicing correct pipetting procedure doesn't have to be boring. "Pipetting by Coordinates" is an effective way to teach necessary pipetting skills in an enjoyable manner. Students create designs as they add volumes of colored water to specific wells and gain experience using a basic biotechnology tool.

Key Words: Micropipetting.
Subject: Pipettes (Usage)
Volumetric analysis (Methods)
Volumetric analysis (Study and teaching)
Author: Bokor, Julie
Pub Date: 08/01/2012
Publication: Name: The American Biology Teacher Publisher: National Association of Biology Teachers Audience: Academic; Professional Format: Magazine/Journal Subject: Biological sciences; Education Copyright: COPYRIGHT 2012 National Association of Biology Teachers ISSN: 0002-7685
Issue: Date: August, 2012 Source Volume: 74 Source Issue: 6
Geographic: Geographic Scope: United States Geographic Code: 1USA United States
Accession Number: 298172406
Full Text: For the vast majority of the population, pipetting isn't a frequently used skill, but for one class period, the fate of a molecular biology experiment is determined by the skilled use of a $300 piece of equipment. Often the learning objective isn't the correct use of a micropipettor, but rather the steps that produce the amplified DNA band on a gel.

Over the past decade, our center has grappled with adequately training secondary science students and teachers to use micropipettes, trying to balance time spent practicing versus time investigating restriction enzymes or solving a crime using DNA gel electrophoresis. Because we often have students on campus for only a few hours, our goal is to provide an educational and enjoyable science experience that will hopefully leave a lasting positive impression.

Through the years, we have used most micropipetting practice procedures: Petri dishes with rows of wells, plastic gels, microfuge tubes, and colored water. When we switched to a bufferless electrophoresis system, we no longer had to worry about students puncturing their gels. The focus really could be on accurately using a micropipette to measure small volumes of liquid. The Battleship method--or "Pipetting by Coordinates"--was born.

Students are given a set of coordinates that correspond to the wells of a 96-well plate. Students use different volumes and colors of water to create a design in the plate. Eight designs are used by students and teachers in our precollege programs, all with success (see Figure 1 for sample design protocol). There are varying levels of protocol difficulty based on the number of volume changes and complexity of the design. All have volumes requiring the use of both a P20 and P200 micropipettor.

For our purposes, this activity is the perfect introduction to using a micropipette. Proper procedure is modeled by the teacher and then corrected individually while checking to make sure that students are only going to the first stop to draw up their sample. The satisfaction of seeing a design materialize (see Figure 2 for an example) is much more gratifying than moving tiny amounts of clear liquid from one tube to the next. Completed plates can be projected for the class to see and discuss. As an additional assessment, students can determine the total volume of liquid they added to the plate and the corresponding mass (1000 [micro]L water = 1 g) and, using a tared scale, determine the actual mass of the completed design. For example, the total volume for the bacteriophage design is 4462 [micro]L = 4.462 g. More than a 1-2% variance indicates a micropipetting error that needs to be investigated further. Possible causes could be going to the second stop before drawing up the sample, or a missed well.

Our micropipetting stations are for student pairs. Each duo receives 50-mL conical tubes with colored water, one P20, one P200, one box of disposable P20-P200 tips, one 96-well plate, and directions. To conserve tips, we ask students to use one tip per micropipettor. The conical tubes, 96-well plate, and directions can all be reused.

The response to this activity has been quite favorable. It is a welcome addition to our biotechnology explorations, with numerous benefits for students and teachers (see below). If you are looking for a way to add a bit of excitement to your micropipetting routine, have your students give this activity a try. The Pipetting by Coordinates lesson plan and additional design protocols are available from the author or online at http://www.cpet.ufl.edu.

* Benefits of Pipetting by Coordinates

1. Student and teacher interest and engagement are higher than with other practice methods.

2. Students have to change volumes multiple times, allowing them to practice adjusting the volume and observing the different volumes in the tip and plate.

3. The activity takes ~20 minutes, allowing time to observe each student and correct micropipetting errors (designs and volumes can be adjusted to reduce time required.)

4. Increased confidence using molecular biology equipment and with laboratory skills.

5. Students can create their own designs unique to the season or school. Ours are particular to our university and content covered in programs.

6. Assessment is as quick as verifying that the image is correct or determining the mass of the final product.

[FIGURE 2 OMITTED]

DOI: 10.1525/abt.2012.74.6.11

JULIE BOKOR is Assistant Director of the Center for Precollegiate Education and Training at the university of Florida, 331 Yon Hall, Box 112010, Gainesville, FL 32611. e-mail: julie@cpet.ufl.edu.
Figure 1. Bacteriophage design protocol.

Protocol E:

Micropipette indicated amounts into designated
wells on the 96 well plate.

Using the RED dye,
20 [micro]L: A12
16 [micro]L: C3, C4
17 [micro]L: D2, D3, D4, D5, D12
18 [micro]L: E3, E4
19 [micro]L: G12

Using the PURPLE dye,

8 [micro]L: A10, A11, B3, B4, B9
6 [micro]L: D1, D6, D7, D8, D9, D10, D11
8 [micro]L: E2, E5, E9
9 [micro]L: F3, F4, F9
7 [micro]L: C2, C5, C9, G10, G11

Using the RED dye,
70 [micro]L: A12
116 [micro]L: C3, C4
110 [micro]L: D2, D3, D4, D5, D12
93 [micro]L: E3, E4
85 [micro]L: G12

Using the PURPLE dye,
118 [micro]L: A10, A11, B3, B4,B9
133 [micro]L: D1, D6, D7, D8, D9, D10, D11
122 [micro]L: E2, E5, E9
129 [micro]L: F3, F4, F9
141 [micro]L: C2, C5, C9, G10, G11
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