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_x000D_

• Describe the morphology of plaques caused by different bacteriophages
_x000D_
• Observe how small mutations in the genome of a bacteriophage can change the progression of its infection of its host bacteria
_x000D_
• Practice sterile technique and how to perform serial dilutions
_x000D_
• Discuss the importance of controls
_x000D_
• Draw conclusions from observed data
_x000D_
• Demonstrate the understanding that patterns seen in simpler biological systems can help in the understanding of the biological systems of more complex organisms
_x000D_
• 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_x000D_

• Familiarity with sterile technique
_x000D_
• Basic knowledge of bacteria, DNA, and proteins

Kit ships with perishable materials included. Note: Refrigerate coliphage until use; do not freeze. Keep bacteria culture and agar at room temperature.

For Educational Use Only. All orders are subject to Carolina’s Terms and Conditions

Name	Include Note
Bunsen Burner	Needed, Not Included
Test Tubes	Included
Tryptone Base Layer Agar Bottles	Included
Tryptone Soft Agar, 50 mL	Included
Serial Dilution Tryptone Broth Tubes, 9 mL	Included
Carolina Science Online® Resources (1-year subscription)	Included
Sterile Serological Pipets, 10 mL	Included
Bulb Pipets, 1 mL	Included
Teacher's Manual with Reproducible Student Guide	Included
Pipetting Devices for Serological Pipets	Needed, Not Included
Paper Towels	Needed, Not Included
Escherichia coli B Agar Slant Culture	Included
Coliphage T4 (104 particles/mL) Vial	Included
Sterile Serological Pipets, 5 mL	Included
Sterile Serological Pipets, 1 mL	Included
Petri Dishes	Included
Tryptone Broth Tubes, 5 mL	Included
Permanent Markers	Needed, Not Included
Coliphage T4r (104 particles/mL) Vial	Included
Water Bath, 48° C	Needed, Not Included
Test Tube Racks	Needed, Not Included
Incubator, 37° C	Needed, Not Included
Microwave Oven or Boiling Water Bath	Needed, Not Included
Thermometer	Needed, Not Included