An investigative graduate laboratory course for teaching modern DNA techniques

Abstract

This graduate-level DNA methods laboratory course is designed to model a discovery-based research project and engages students in both traditional DNA analysis methods and modern recombinant DNA cloning techniques. In the first part of the course, students clone the Drosophila ortholog of a human disease gene of their choosing using Gateway® cloning. In the second part of the course, students examine the expression of their gene of interest in human cell lines by reverse transcription PCR and learn how to analyze data from quantitative reverse transcription PCR (qRT-PCR) experiments. The adaptability of the Gateway® cloning system is ideally suited for students to design and create different types of expression constructs to achieve a particular experimental goal (e.g. protein purification, expression in cell culture, and subcellular localization), and the genes chosen can be aligned to the research interests of the instructor and/or ongoing research in a department. Student evaluations indicate that the course fostered a genuine excitement for research and in depth knowledge of both the techniques performed and the theory behind them. Our long-term goal is to incorporate this DNA methods laboratory as the foundation for an integrated laboratory sequence for the Master of Science degree program in Molecular and Cellular Biology at Quinnipiac University, where students use the reagents and concepts they developed in this course in subsequent laboratory courses, including a protein methods and cell culture laboratory.

The Impact of Innovative Teaching Approaches on Biotechnology Knowledge and Laboratory Experiences of Science Teachers

“…Similar results have been obtained in the studies related to the applied teaching of biotechnology subjects in the literature. It was stated that the discovery-based laboratory course using the DNA analysis methods and recombinant DNA cloning techniques enabled the students to gain detailed information about the laboratory techniques used [59]. Studies on the use of DNA technologies (PCR, electrophoresis, etc.)…”

Evaluating two steps in transcription using a fluorescence‐based electrophoretic mobility shift assay

“…This complements, but is distinct, from other experiments in the education literature that probe transcription by evaluating changes in the expression level of reporter genes such as betagalactosidase, [15][16][17] or evaluating levels of cellular RNA via RT-PCR or arrays, for example. [18][19][20][21] This experiment is appropriate for students in upperlevel undergraduate biochemistry or molecular biology laboratory courses. It is not necessary that students have been introduced to EMSAs or fluorescence, although it is helpful if they have been introduced to transcription and the central dogma in classroom coursework.…”

“…This experiment provides an experimental framework for students to explore the mechanism of transcription by distinguishing between the binding of RNAP and its transcriptional activity. This complements, but is distinct, from other experiments in the education literature that probe transcription by evaluating changes in the expression level of reporter genes such as beta‐galactosidase, 15–17 or evaluating levels of cellular RNA via RT‐PCR or arrays, for example 18–21 …”

Expression, Purification and Characterization of RecJ Nuclease -An Integrated Biochemistry Laboratory Course for Undergraduates

“…At the same time, it is difficult for students to apply the techniques learned in the experimental class to their own future research projects flexibly. Until now, many schools are always trying to optimize the teaching mode of the experimental course [1][2][3][4][5][6]. Teachers also tried to improve the curriculum mode in the integrated biochemistry experiment.…”