Description
Join a nationwide monitoring program that tests for antibiotic resistance genes in environmental soil samples. Students will collect soil samples based on their own hypotheses about antibiotic resistance hotspots, extract total environmental DNA from soil, and use the molecular methods of PCR and gel electrophoresis to test their samples for evidence of tetracycline resistance. They will then contribute their data to a national database of antibiotic resistance surveillance.
This lab has been developed in conjunction with the PARE (Prevalence of Antibiotic Resistance in the Environment) project. The PARE project engages students to test and report the prevalence of tetracycline-resistant bacteria from soil at diverse geographic sites, engaging students in one of the great environmental and health challenges of our time. Learn more about the PARE project: https://sites.tufts.edu/ctse/pare
- Techniques: Micropipetting, eDNA extraction, PCR, gel electrophoresis
- Topics: Environmental monitoring, molecular biology, biotechnology
- Time required: This is a project-based activity that requires at least three to four class periods for completion
- Level: Advanced high school, college
This is an advanced lab activity with unknown outcomes and is ideal for independent research projects. Students performing this lab should have experience following complex molecular protocols. For a case study approach to the same curriculum, try the Antibiotic Resistance Lab.
This lab kit provides enough reagents for 5 lab groups:
- ZymoBIOMICS DNA Miniprep Kit
- EZ PCR Master Mix, Load-Ready™
- 16S Primer Mix
- tetB Primer Mix
- tetM Primer Mix
- Positive Control Plasmid
- Fast DNA Ladder 1
- Nuclease Free Water
Supplied by user:
Standard reagents and consumables must be supplied by the user. The Learning Lab Companion Kit (catalog no. KT-1510-01) contains all the consumables (except water and 95% ethanol) needed for 5 or more lab groups. These reagents and consumables can also be sourced independently.
Reagents and consumables supplied by user:
- Agarose (electrophoresis grade)
- DNA stain (e.g., SeeGreen™ or GelGreen®)
- Gel electrophoresis buffer (e.g., 1X TBE)
- Distilled or deionized H2O (to dilute buffer concentrate – not included in Lab Companion Kit)
- Microtubes
- PCR tubes
Reagents not included in Lab Companion Kit:
- 95% ethanol
Required equipment:
- Micropipettes and tips
- Tissue homogeneizer or vortex mixer with microtube holder attachment
- Microcentrifuge
- Thermal cycler
- Gel electrophoresis and visualization system
Storage notes:
- Reagents may ship at room temperature, but require freezer storage upon arrival
- Frozen reagents are stable for 12 months when stored in freezer
- ZymoBIOMICS DNA Miniprep Kit is stable for 6 months when stored at room temperature
Downloads
- miniPCR eDNA Project Teacher’s Guide
- miniPCR eDNA Project Student’s Guide
- miniPCR eDNA Project Classroom Slides
- SDS – EZ PCR Master Mix
- SDS – Fast DNA Ladder 1
- SDS – Primer Mix
- SDS – Positive Control DNA
- SDS – ZymoBIOMICS DNA Miniprep Kit
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