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 using PCR and 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 8 lab groups of 4 students with each group running the four PCR reactions 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:
Free miniPCR Learning Lab™ downloads:
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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