Basic Methods In Molecular Biology

The new edition of this popular book emphasizes the decisions that need to be made to select one procedure over another.

This edition emphasizes the decisions that need to be made to select one procedure over another in the everyday workings of a molecular biology laboratory. Trouble-shooting problems that may emerge when performing such methods are included and concise instructions are provided.

Basic Methods in Molecular Biology discusses the heart of the most recent revolution in biology—the development of the technology of genetics. The achievements in this field have simply changed what biologists do and, perhaps even more important, the way they think. Moreover, never before have scientists from such a broad range of disciplines rushed into such a small and slightly arcane field to learn and carry off a bit of the technology. This book comprises 21 chapters, opening with three introductory ones that discuss the basics of molecular biology; the tools of the molecular biologist; and general preparations, procedures, and considerations for use of the book. The following chapters then discuss cloning vectors and bacterial cells; preparation of DNA from eukaryotic cells; probing nucleic acids; plasmid DNA preparation; DNA restriction fragment preparation; purification of DNA; and preparation and analysis of RNA from eukaryotic cells. Other chapters cover preparation of DNA from bacteriophage clones; cloning DNA from the eukaryotic genome; subcloning into plasmids; M13 cloning and sequencing; further characterization of cloned DNA; transfection of mammalian cells in culture; protein methods; general methods; and specialized methods. This book will be of interest to practitioners in the fields of biology and molecular genetics.

This laboratory manual gives a thorough introduction to basic techniques. It is the result of practical experience, with each protocol having been used extensively in undergraduate courses or tested in the authors laboratory. In addition to detailed protocols and practical notes, each technique includes an overview of its general importance, the time and expense involved in its application and a description of the theoretical mechanisms of each step. This enables users to design their own modifications or to adapt the method to different systems. Surzycki has been holding undergraduate courses and workshops for many years, during which time he has extensively modified and refined the techniques described here.

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 third 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

Advanced Methods in Molecular Biology and Biotechnology: A Practical Lab Manual is a concise reference on common protocols and techniques for advanced molecular biology and biotechnology experimentation. Each chapter focuses on a different method, providing an overview before delving deeper into the procedure in a step-by-step approach. Techniques covered include genomic DNA extraction using cetyl trimethylammonium bromide (CTAB) and chloroform extraction, chromatographic techniques, ELISA, hybridization, gel electrophoresis, dot blot analysis and methods for studying polymerase chain reactions. Laboratory protocols and standard operating procedures for key equipment are also discussed, providing an instructive overview for lab work. This practical guide focuses on the latest advances and innovations in methods for molecular biology and biotechnology investigation, helping researchers and practitioners enhance and advance their own methodologies and take their work to the next level. Explores a wide range of advanced methods that can be applied by researchers in molecular biology and biotechnology Features clear, step-by-step instruction for applying the techniques covered Offers an introduction to laboratory protocols and recommendations for best practice when conducting experimental work, including standard operating procedures for key equipment

This manual is designed as an intensive introduction to the various tools of molecular biology. It introduces all the basic methods of molecular biology including cloning, PCR, Southern (DNA) blotting, Northern (RNA) blotting, Western blotting, DNA sequencing, oligo-directed mutagenesis, and protein expression. Provides well-tested experimental protocols for each technique Lists the reagents and preparation of each experiment separately Contains a complete schedule of experiments and the preparation required Includes study questions at the end of each chapter

This volume has evolved from a laboratory methods book that one of us first compiled nearly fifteen years ago. Since that time the book has undergone many minor revisions in order to include new methods and updated versions of older methods. The result has been an increasingly useful and more widely circulated book. However, the recent series of technological explosions generally lumped together under the name of "recombinant DNA technology" has been a turning point in the evolution of this previously underground publication. Minor revisions will no longer do. To keep the book useful we have had to make major revi sions and additions. The result is a dramatically expanded book that should be more useful to more people. The larger size and wider usefulness of the book have made this more formal publication seem a reasonable step to take. One of the reasons that this volume should be useful to many people is that it includes only procedures that have been used repeatedly by us and that have proven highly reliable both to ourselves and to others in our laboratories.