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

Manipulation and Expression of Recombinant DNA: A Classroom Laboratory Manual

Science & Math

Carson, Sue

Robertson, Dominique

Biological Sciences

Academic Press

This laboratory manual will be an indispensable tool for students, by providing in depth descriptions of protocols which cover the basic aspects of molecular biology and biochemistry. The book covers techniques in detail, and teaches students to culture and transform bacteria, grow vector plasmids, purify insert DNA, ligate vector and insert, and ultimately express protein and assay its activity. Comprehensive and complete, this text covers the underlying techniques used in all basic research and biotechnology laboratories. Key Features* Designed to cover the basic aspects of molecular biology and biochemistry* Features student-tested protocols* Presents experiments in a modular fashion, lending itself equally to the traditional semester course and the shorter intensive course.* Covers important techniques and protocols used in all basic research labs* "Project" approach to experiments gives students an overview of entire process* Experiments incorporate immunological techniques that go beyond basic exercises in DNA cloning

Manipulation and Expression of Recombinant DNA: A Laboratory Manual

Miller, David M.

Shore, Scott

On the forefront of modern scientific innovation, Cloning, Gene Expression and Protein Purification: Experimental Procedures and Process Rationale effectively doubles as a laboratory manual for students and a reference book for professional researchers. Designed for advanced undergraduate and beginning graduate students in molecular biology, this unique combination lecture/laboratory resource presents detailed protocols for the multi-step process involved in isolating a gene, cloning and characterizing it, expressing its encoded protein, and purifying and characterizing the protein's basic physical properties. This manageable volume includes both theoretical background and practical procedures and is structured around twenty experiments that demonstrate how to prepare, manipulate, and analyze plasmids, produce fusion proteins in bacteria, and purify these proteins based on unique chemical properties or substrate affinities. The book describes advanced topics such as the use of antibodies and the techniques developed to transform their structures, as well as combinatorial approaches designed to manipulate the structure and functions of proteins and nucleic acids. Supplemental literature provides a variety of theoretical explanations encouraging a more intuitive understanding of the experimental mechanisms and behaviors of the chemical participants, while also giving students the tools needed to become "capable proactive researchers." Features: � Emphasizes electrophoresis, Southern and Western blotting, and combinatorial techniques � Defines clear reaction mechanisms; stipulates the functions of reagents; and helps students think about the precise consequences of solution and procedural manipulations � Discusses fluorophores, and solvent effects on protein structure � Characterizes plasmids, cDNAs, and antibody probes (available from ATCC) in research literature � Includes carefully selected primary source research literature and articles from current vendor literature � Contains a glossary of unfamiliar phrases and jargon; important summary statements and conclusions are italicized � Provides an alphabetized list of common reagents for rapid reference � Offers an extensive index of concepts and terms � Categorizes helpful and distinctive information into five types of supplemental literature: Innovation/ Insight, Theory/Principle, Process Rationale, Vendor Literature, and Alternative Approaches

Cloning, Gene Expression, and Protein Purification: Experimental Procedures and Process Rationale

MILLER, William

Hardin, Charles

Edwards, Jennifer

Riell, Andrew

Presutti, David

Oxford University Press

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