Culture-Based Modules

For a culture-based approach to surveillance, students start with the Core PARE Module in which soil is tested for the prevalence of live bacteria that are resistant to tetracycline and upload results to our database. From there, students in biosafety level 2-equipped laboratories can sub-culture resistant colonies and do further tests.


Colony Identification

16S PCR from colony

Students choose a colony isolated from the core PARE module for tentative phylogenetic characterization. Genomic DNA is isolated and used in PCR reactions to amplify the 16S rRNA gene. Amplification results are confirmed by gel electrophoresis, the amplified DNA cleaned and sent for sequencing. Sequences are compared to those in the Ribosomal Database Project database to assign putative identity. Identity information is a critical early step to identify potential pathogens so this module is recommended prior to any future modules that require tetracycline-resistant colony sub-culturing.


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Acknowledgements

This module was developed in collaboration with Dr. Brad Goodner, Hiram College.

NOTE: This activity is BSL-2. Students will be handling environmental unknowns. Please use the “request access” page prior to embarking on this project.

Multi-drug Resistance

Horizontal Transfer