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This lab introduces students to bacteriophages and how a simple mutation in the phage genome can change the progression of the phage's infection of a bacterial population. Students will use the double-agar layer technique to plate the T4 and T4r mutant coliphage, as well as a control plate. The next day they will count and describe the morphology of the plaques created by each of the 2 different types of bacteriophage.

This kit demonstrates the effect a single mutation can have on the progression of a viral disease. Students create serial dilutions of 2 strains of the T4 coliphage (T4 and T4r). They infect a host culture of E. coli B and plate the infected cultures using an agar-layer technique. By looking at the difference in plaque morphology between the plates infected with the 2 strains of coliphage, students observe how small mutations in the genome of a bacteriophage can change the progression of its infection of its host bacteria. Students also learn and practice several techniques invaluable in microbiology, including sterile technique, serial dilution, titer calculation, and agar-layering.

Materials are suitable for 1 to 2 groups of students working in groups of 4. The hands-on portion of the lab can be completed in one 40-minute session followed by a 15-minute session the next day. The prelab can be completed in 30 minutes, and the analysis and assessment can be completed in 30 minutes.

This kit replaces and extends the Plaque Demonstration Kit A (items #124310 and #124310P).

Objectives

• Describe the morphology of plaques caused by different bacteriophages
• Observe how small mutations in the genome of a bacteriophage can change the progression of its infection of its host bacteria
• Practice sterile technique and how to perform serial dilutions
• Discuss the importance of controls
• Draw conclusions from observed data
• Demonstrate the understanding that patterns seen in simpler biological systems can help in the understanding of the biological systems of more complex organisms
• Use knowledge of the bacteriophage lytic life cycle to answer questions about the effect of bacteriophages on the bacterial host and its environment

Prerequisite Knowledge and Skills

• Familiarity with sterile technique
• Basic knowledge of bacteria, DNA, and proteins

Includes a voucher to request perishable materials later at your convenience. Contact us or return the voucher to request delivery of perishables. Note: Refrigerate coliphage until use; do not freeze. Keep bacteria culture and agar at room temperature.

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