Antibiotic-resistant bacteria have been traced to one farm and neighbours are concerned. Use PCR to test soil samples from neighbouring farms to determine if the antibiotic-resistant bacteria are present.
This lab (formerly known as “miniPCR® Antibiotic Resistance Lab: Monitoring resistant organisms in the environment“) presents a fictional case study of a very real problem. Epidemiologists have traced an outbreak of antibiotic-resistant bacteria to one farm. Students will use PCR to determine if the antibiotic-resistant bacteria have spread to neighbouring areas. This case study introduces students to how molecular analyses can serve as a tool in environmental monitoring and surveillance. Produced in collaboration with PARE – The Prevalence of Antibiotic-Resistance in the Environment project at Tufts University.
No harmful samples are used in this lab.
Key info and benefits:
- Techniques: Micropipetting, PCR, gel electrophoresis
- Time required: 90 minutes - can be done in two separate lessons or one double lesson
- Level: High school
- This kit provides enough reagents for eight groups (32 students).
- These reagents are optimised for blueGel™ electrophoresis systems. For post-staining techniques, more sample may be required per well.
- 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.
Kit includes:
- 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
Notes:
- 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.
