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

Area of Science:

• Microbiology
• Molecular Biology
• Bioinformatics

Background:

• The COVID-19 pandemic necessitated a shift to online laboratory activities for microbiology.
• Introductory students often grasp basic PCR principles but lack understanding of specificity and error generation in diagnostic tests.

Purpose of the Study:

• To develop a virtual Polymerase Chain Reaction (PCR) primer design and activity for introductory students.
• To enhance understanding of molecular biology concepts, PCR specificity, and diagnostic test limitations within the context of COVID-19 testing.

Main Methods:

• A simple PCR primer design and virtual PCR activity were created for online or traditional lab settings.
• The activity was designed to reinforce molecular biology concepts relevant to pandemic-era diagnostics.

Main Results:

• Nursing, Biology, and non-Biology majors successfully completed the virtual worksheet with minimal instructor assistance.
• The activity effectively engaged students in reviewing molecular biology principles.

Conclusions:

• The virtual PCR activity is a versatile educational tool for introductory microbiology and molecular biology.
• It can be implemented as a standalone module, a warm-up for bioinformatics, or a pre-lab exercise for quantitative PCR.
• The activity can be adapted for both introductory and advanced students with minor modifications.