title: 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/or 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. © 2017 by The International Union of Biochemistry and Molecular Biology, 45(4):351-359, 2017.
authors.affiliation: Department of Biological Sciences, Quinnipiac University, Hamden, Connecticut, 06518.
authors.forename: Alexandre
authors.initials: A
authors.lastname: de Lencastre
authors.affiliation: Department of Biological Sciences, Quinnipiac University, Hamden, Connecticut, 06518.
authors.forename: A
authors.initials: A
authors.lastname: Thomas Torello
authors.affiliation: Department of Biological Sciences, Quinnipiac University, Hamden, Connecticut, 06518.
authors.forename: Lani C
authors.initials: LC
authors.lastname: Keller
mesh_terms: D000818:Animals; D001671:Biochemistry / Q000193:education*; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods*; D003479:Curriculum; D004268:DNA-Binding Proteins / Q000235:genetics*; D029721:Drosophila Proteins / Q000235:genetics*; D004331:Drosophila melanogaster / Q000235:genetics; D004500:Education, Graduate / Q000379:methods; D004521:Educational Measurement / Q000706:statistics & numerical data; D015870:Gene Expression; D006801:Humans; D008967:Molecular Biology / Q000193:education*; D010750:Phosphoproteins / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D017422:Sequence Analysis, DNA; D013334:Students; D014169:Transformation, Bacterial
chemical_list: D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D010750:Phosphoproteins; C077938:cact protein, Drosophila
keywords: DNA techniques; Gateway® system; Integration of research into undergraduate teaching; cloning; laboratory exercises; molecular biology; new course development