Manipulation and Expression of Recombinant DNA

This manual is an indispensable tool for introducing advanced undergraduates and beginning graduate students to the techniques of recombinant DNA technology, or gene cloning and expression. The techniques used in basic research and biotechnology laboratories are covered in detail. Students gain hands-on experience from start to finish in subcloning a gene into an expression vector, through purification of the recombinant protein.The second edition has been completely re-written, with new laboratory exercises and all new illustrations and text, designed for a typical 15-week semester, rather than a 4-week intensive course. The "project approach to experiments was maintained: students still follow a cloning project through to completion, culminating in the purification of recombinant protein. It takes advantage of the enhanced green fluorescent protein-students can actually visualize positive clones following IPTG induction.*Cover basic concepts and techniques used in molecular biology research labs*Student-tested labs proven successful in a real classroom laboratories*Exercises simulate a cloning project that would be performed in a real research lab*"Project" approach to experiments gives students an overview of the entire process*Prep-list appendix contains necessary recipes and catalog numbers, providing staff with detailed instructions

Dr. Susan Carson is a Professor of Plant and Microbial Biology at North Carolina State University (Raleigh, NC) where she has served on the faculty since 2001. She directs the Master of Microbial Biotechnology Program and also leads a uni?versity-wide faculty development program focused on enhancing students' critical and creative thinking skills across disci?plines. Prior to her current role, Dr. Carson spent over a decade leading curriculum development for the North Carolina State Biotechnology Program and two years as a Program Officer at the National Science Foundation Division of Undergraduate Education. Her current work focuses on college-level biology education, enhancing students' higher order thinking across disciplines, and integration of Design Thinking in a Professional Science Masters (PSM) program. She graduated from Rutgers University (New Brunswick, NJ) with a BS in Biotechnology, and from the University of North Carolina (Chapel Hill, NC) with a PhD in Microbiology.

ForewordAcknowledgementsNote to InstructorsInstrumentationNomenclatureINTRODUCTION: Conceptual outline for experimentsPART I MANIPULATION OF DNAPART II SCREENING TRANSFORMANTSPART III EXPRESSION, DETECTION, AND PURIFICATION OF RECOMBINANT PROTEINS Appendices: Equipment, Prep list, Making sense of orientation