Primer Design for Reverse Transcription-Quantitative PCR Simulation: an Online Lab Exercise Using a SARS-CoV-2 Model

Since the start of the 2019 coronavirus disease (COVID-19) pandemic, many microbiology lab activities have been conducted online. We produced a simple PCR primer design and virtual PCR activity for introductory students to use as part of an online laboratory course or as an independent activity in a traditional laboratory setting. Most students are aware of basic PCR concepts but struggle with important details, such as how PCR is specific and how false positives and negatives can be generated in a diagnostic test that is not well designed. This exercise helps students review molecular biology concepts within the context of a test that was commonplace during the COVID-19 pandemic. We found that nursing students and Biology and non-Biology majors were able to complete the worksheet as a group with minimal instructor input. This could be used as a stand-alone activity, as a warm-up for other bioinformatics exercises, or as a prelab activity for actual in-lab quantitative PCR experiments, such as the one offered by miniPCR bio. With minor modifications, it could also be used with more advanced students.