Presents techniques tested at the Curie Institute and other leading labs and lists all commercially available enzymes, vectors, linkers, and other basic products for ready reference. Offers detailed explanation of protocols, allowing the isolation, cloning, and expression of genes from living species. Presents up-to-date techniques on sequencing, in vitro expression of cloned gene, and use of computers for study of nucleic acids, and is the only book that shows how to isolate DNA-protein complexes and new methods for mutagenesis of cloned genes. Contains 235 figures and 80 tables.

This comprehensive guide to gene cloning provides beginning and advanced readers with the background, standard techniques, practical applications, and ethical and safety considerations in the field. A one-stop reference for students, researchers in academia and industry, and anyone interested in a thorough but accessible overview.

The two Essential Molecular Biology books in the Practical Approach Series are designed for the absolute beginner at gene cloning whether they be at the start of their career or an experienced researcher in another field. As with the first editions, the objective of both volumes is to combinesolid practical information with sufficient background material to ensure that the novice can understand how a technique works, what it achieves, and how to make modifications to suit personal requirements.Volume 2 details procedures for isolating and studying individual genes (preparation and screening of libraries, polymerase chain reactions, DNA sequencing and studying gene expression). It is assumed that the basics of Volume 1 are now in place, but the procedures are still described in the samedown to earth fashion with protocols complemented by background information and troubleshooting hints.

This textbook introduces readers in an accessible and engaging way to the nuts and bolts of protein expression and engineering. Various case studies illustrate each step from the early sequence searches in online databases over plasmid design and molecular cloning techniques to protein purification and characterization. Furthermore, readers are provided with practical tips to successfully pursue a career as a protein engineer. With protein engineering being a fundamental technique in almost all molecular biology labs, the book targets advanced undergraduates and graduate students working in molecular biology, biotechnology and related scientific fields.

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

This textbook introduces readers in an accessible and engaging way to the nuts and bolts of protein expression and engineering. Various case studies illustrate each step from the early sequence searches in online databases over plasmid design and molecular cloning techniques to protein purification and characterization. Furthermore, readers are provided with practical tips to successfully pursue a career as a protein engineer. With protein engineering being a fundamental technique in almost all molecular biology labs, the book targets advanced undergraduates and graduate students working in molecular biology, biotechnology and related scientific fields.

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

Chromosome Microdissection and Cloning: A Practical Guide is a straightforward guide to chromosome microdissection and cloning. It presents an overview of the procedures and briefly reviews a few areas of research in which these techniques are applied. Topics range from preparation of chromosomes for microdissection to molecular cloning of microdissected chromosomal DNA. Methods of chromosome microdissection, including video microscope method and oil chamber method, are described. Comprised of five chapters, this book begins with an overview of the structure and organization of chromosomes, followed by a description of methods for preparing and preserving chromosomal DNA in a manner that is useful for cloning and direct analysis. Microdissection of metaphase chromosomes and isolation of fragments can be accomplished in one of the three ways described in the next chapter: by microdissection using an upright microscope and glass capillaries in an oil chamber; by laser microbeam; and with the use of an inverted microscope equipped with a video camera and high magnification-high resolution lenses. A step-by-step guide to these techniques and solutions for common problems are given following each method. Protocols for cloning and identifying genetic sequences from defined chromosome regions, particularly using the polymerase chain reaction, are also discussed. The final chapter focuses on applications of chromosome microdissection, such as cloning of disease-specific genes and generating "sequence tagged sites" to be used in large DNA sequencing projects. This monograph will be particularly helpful to investigators setting up microdissection systems de novo.