miniPCR™ Antibiotic Resistance Lab:Monitoring resistant organisms in the environment

Description

Produced in collaboration with PARE –The Prevalence of Antibiotic-Resistance in the Environment project at Tufts University.

Students are presented with information regarding a fictitious outbreak of food-borne infection that is resistant to treatment with the antibiotic carbapenem and carries the NDM-1 gene (encoding an enzyme that hydrolyzes carbapenems.) Epidemiologists have identified the source of the outbreak, but neighboring farms are worried. Students are tasked with usingPCRand gel electrophoresis to establish whether farms are at risk from antibiotic-resistant bacteria that may be spreading in the environment.

This known-outcome molecular-based case study introduces students to the problem of environmental surveillance for antibiotic resistance genes. We have designed this activity in collaboration with PARE to give students a robust, easily reproducible simplified environmental microbiology scenario which can serve as an introduction to authentic environmental monitoring research. No potentially harmful environmental samples are used in this lab.

• Techniques utilized: Polymerase Chain Reaction (PCR), DNA gel electrophoresis.
• Time required: can be completed in two 45 min. class periods.
• Reagents needed: ‘miniPCR™ Antibiotic Resistance Lab’ reagents kit (included in this product), electrophoresis reagents (available separately)

Suggested skill level: Intended for any student seeking familiarity with PCR, DNA gel electrophoresis, environmental microbiology and antibiotic resistance, from middle school to college.

The miniPCR Antibiotic Resistance Lab kit contains reagents for 8 lab groups of up to 4 students each (32 students):

• All molecular reagents needed (template DNA, primers, master mix containing dNTPs, polymerase, loading dye) and DNA ladder.
• Enough for 8labgroups of 4 students with each group running the fourPCRreactions of the case study narrative (two test sites two controls).
• Reagents are compatible with miniPCR™,blueGel™or other standard PCR and DNA electrophoresis systems

Please note:

• These reagents require frozen storage.
• Microtubesand electrophoresis reagents sold separately.

Free miniPCR Learning Lab™downloads:

• Antibiotic Resistance Lab Instructor’s Guide
• Antibiotic Resistance Lab Student’s Guide
• Antibiotic Resistance Lab Classroom Slides

Optional: Lab Companion Kit

Microtubes (50), PCR tubes (100), agarose (10 blueGel Tabs x 0.4 g ea.), 20X TBE buffer (30 mL), GelGreen™ DNA stain, 10,000X (20 µL).

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