Use PCR to determine trace the spread of antibiotic-resistant bacteria!
Students are presented with information regarding a fictitious outbreak of a food-borne infection that is resistant to treatment with the antibiotic carbapenem and carries the NDM-1 gene (encoding an enzyme that hydrolyses carbapenems.) Epidemiologists have identified the source of the outbreak, but neighbouring 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.
Key info and benefits:
- Techniques: Micropipetting, PCR, gel electrophoresis
- Time required: 90 minutes
- Level: High school
- This kit provides enough reagents for eight groups (32 students). These reagents are compatible with blueGel™ or other standard DNA electrophoresis systems.
- Introduces students to the problem of environmental surveillance for antibiotic resistance genes.
- Designed 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.
- Master mix containing dNTPs, polymerase and loading dye (240uL)
- Primer mix (480uL)
- DNA samples (150uL each)
- DNA controls (150uL each)
- DNA ladder with loading dye (100uL)
- See notes for extra requirements
- See resources below to download the teacher's guide and student guide for this lab. Contact team @ Southern Biological for answers key.
- This kit provides DNA samples, DNA ladder and gel loading dye only. To perform this lab, you will require PCR and electrophoresis equipment; such as electrophoresis chamber, agarose, TBE buffer and Midori green gel stain. A miniPCR unit for eight or sixteen tubes, PCR tubes and our electrophoresis starter kit contains the hardware and reagents required.
- For best results, we recommend running your gels in the blueGel electrophoresis chamber. If you do not have a blueGel electrophoresis unit with built-in illuminator, we have a transilluminator onto which you can place your gels to visualise your results immediately after the electrophoresis run is complete.
- These DNA samples require frozen storage, which will keep them stable for 12 months. The DNA samples are stable for a couple of weeks at room temperature, and the master mix for several days.
- No potentially harmful environmental samples are used in this lab.