Thomas a Mennella

Making Microtubules Glow Thomas a Mennella

* For more in-depth information and resources, visit this manual's website: http: //thomasmennella.wix.com/mtglow * The importance of a robust undergraduate research experience has been demonstrated time and again. However, too few undergraduates engage in genuine research and leverage this opportunity. This laboratory manual is intended to accompany a laboratory course in Cell and/or Molecular Biology that is designed to mimic a true research project. Students work through a 10-step experimental design culminating in the construction, expression, and visualization of microtubules fused to green fluorescent protein in baker's yeast. The steps of this project include the isolation of the tubulin gene (TUB1) from yeast genomic DNA, the cloning of that gene into an expression vector, the amplification of this plasmid in E. coli, and the validation of expression of fluorescent tubulin in yeast via western blot. The semester ends with the visualization of glowing yeast cells by using fluorescent microscopy. Controls and validation steps are embedded throughout the project, as they would be in a genuine research project. This laboratory course more closely resembles a one-semester undergraduate research experience than a typical lab course. However, because courses reach a much larger number of students compared to undergraduate research opportunities, this approach provides students with a valuable research experience that remains confined to the scheduled time block of a typical lab course. With detailed, step-by-step protocols for students to follow (which include the rationale and explanation for key steps), Reflection Questions at the end of each exercise to promote deeper thinking, and thorough Instructor's Notes for each exercise to guide the course instructor through set-up for the day, this manual is easily adopted, and adaptable, for almost any college or university. This lab manual is the companion text for the laboratory course design described in: "Designing Authentic Undergraduate Research Experiences in a Single-Semester Lab Course" published by The American Biology Teacher, Vol. 77 No. 7, September 2015