An adaptable dry lab for SYBR based RT-qPCR primer design to reinforce concepts in molecular biology and nucleic acids.

Abstract

The real time PCR (qPCR) method provides a powerful method to assess levels of particular species of DNA. When combined with reverse transcription (RT-qPCR) it is the predominate technique to measure expression of gene transcripts. While this approach is very powerful, particular care must be taken in the design of the primers to facilitate specific and sensitive detection. Herein describes the framework for an undergraduate assignment which aims to teach primer design for SYBR based RT-qPCR. Beyond gaining direct experience with primer design, students will gain familiarity with important bioinformatic resources as well as a deeper theoretical understanding of the RT-qPCR approach and potential limitations. Moreover, as students' progress through the assignment they re-encounter many important concepts in molecular biology, gene expression, and nucleic acids, creating an opportunity for spiral learning. As this exercise only requires access to free web-based resources and does not require a laboratory it can be used in most science education settings. Despite not being a wet lab, this is a highly authentic research experience as this design process is commonplace in a molecular biology laboratory. Furthermore, the assignment is highly adaptable for different learning outcomes, time frames, and student background and ability. This article seeks to highlight connections and expanded learning outcomes for those already teaching such material, as well as a step-by-step guide for those new to teaching such content.