Utilizing loop-mediated isothermal amplification to detect the presence of Escherichia coli: An inquiry-driven undergraduate laboratory module.

Abstract

The amplification of nucleic acids is a fundamental tool utilized in various scientific disciplines, including Molecular Biology, Immunology, Microbiology, and Genetics. However, due to the time and technology required for traditional polymerase chain reaction and its derivatives, it is not always possible to include such methodologies in undergraduate laboratory curricula. Loop-mediated isothermal amplification (LAMP), a technology that has become increasingly utilized in a variety of laboratory and field settings during the past two decades, is an alternate method of nucleic acid amplification that is rapid, sensitive, and performed under isothermal conditions. We describe an adaptable, inquiry-driven laboratory module that is focused on the detection of Escherichia coli DNA via LAMP amplification. The main objectives of the module are to introduce students to the principles and protocols of LAMP, to help students develop the ability to apply the scientific method to scientific questions, to guide students as they develop the ability to identify the most appropriate methodology to use in the investigation of scientific questions, and to train students to critically evaluate scientific data, theories, and principles and to articulate their evaluations in both written and oral formats.