Plant Molecular Biology A Laboratory Manual

Covering the whole range of molecular biology techniques - genetic engineering as well as cytogenetics of plants -, each chapter begins with an introduction to the basic approach. followed by detailed methods with easy-to-follow protocols and comprehensive troubleshooting. The first part introduces basic molecular methodology such as DNA extraction, blotting, production of libraries and RNA cloning, while the second part describes analytical approaches, in particular RAPD and RFLP. The manual concludes with a variety of gene transfer techniques and both molecular and cytological analysis. As such, this will be of great use to both the first-timer and the experienced scientist.

The book, “A Laboratory Manual of Plant Biotechnology and Molecular Biology” comprises of workable laboratory protocols for a large number of techniques related to plant biotechnology, genetic engineering and molecular biology. This includes plant cell and tissue culture, callus and suspension culture, anther culture, ovule culture, embryo culture, Cryopreservation, Isolation of Plant protoplasts, Protoplast culture and regeneration, production of somatic hybrids through protoplast fusion, gene transformation using Agrobacterium as vector, direct gene transfer using biolistic gun, Isolation of plant and organells DNA, construction and screening of genomic DNA libraries, Molecular markers like RFLP, RAPD, SCARS and CAPS, DNA sequencing, RNA isolation and northern blotting, Isolation of proteins and western blotting etc. The manual is prepared with the objective to cater the needs of post- graduate students as well as for scientists working in the disciplines of Plant Breeding, Genetics, Botany, Plant physiology, Biochemistry, Plant Biotechnology, Molecular Biology etc. It gives an update on some well established methods and presents reliable protocols.

During the past ten years, great advances have been made in the area of plant molecular biology. Such formerly esoteric techniques as gene transfer and plant regeneration are now routinely performed, making the dissection of regulatory elements of genes a common practice in many laboratories. Along with this new technology has come an almost bewildering array of rapidly changing techniques, often making it difficult for the novice to select and perform the technique most appropriate for answering a given biological question. In 1986, some of us felt that many of these techniques had become routine enough to warrant the publication of a laboratory manual. The manual is designed both for advanced college level laboratory courses and as a 'bench guide' for use in the scientific laboratory. Recognizing the rapidly changing nature of plant molecular biology technology, the editors have designed a laboratory manual that is both easy to use in the laboratory and which will be updated as the techniques change and new technologies are devised. Additional chapters that can replace or be added to this first edition will be published periodically. The editors recognize that many of the techniques described in this manual depend upon specialized plant genetic material, microbial strains, or recombinant plasmids. Those people desiring such material should contact the relevant authors directly. A list of the various contributors to this manual, including their addresses, is included.

During the past ten years, great advances have been made in the area of plant molecular biology. Such formerly esoteric techniques as gene transfer and plant regeneration are now routinely performed, making the dissection of regulatory elements of genes a common practice in many laboratories. Along with this new technology has come an almost bewildering array of rapidly changing techniques, often making it difficult for the novice to select and perform the technique most appropriate for answering a given biological question. In 1986, some of us felt that many of these techniques had become routine enough to warrant the publication of a laboratory manual. The manual is designed both for advanced college level laboratory courses and as a 'bench guide' for use in the scientific laboratory. Recognizing the rapidly changing nature of plant molecular biology technology, the editors have designed a laboratory manual that is both easy to use in the laboratory and which will be updated as the techniques change and new technologies are devised. Additional chapters that can replace or be added to this first edition will be published periodically. The editors recognize that many of the techniques described in this manual depend upon specialized plant genetic material, microbial strains, or recombinant plasmids. Those people desiring such material should contact the relevant authors directly. A list of the various contributors to this manual, including their addresses, is included.

Five years ago, the first edition of the Plant Molecular Biology Manual appeared. At that time, the editors felt that the field of plant molecular biology had matured to a point that the publication of a series of protocols in plant molecular biology was warranted. During the past five years, the field of plant molecular biology has expanded rapidly. This expansion is, among other things, reflected by the presence of several journals in the plant sciences, as well as by the increasing amount of plant sciences articles that are published in the more general journals. In 1991 approximately 3000 people attended the Third International Congress of Plant Molecular Biology in Tucson, Arizona, where more than 2000 posters were presented. It is also remarkable to see that nowadays botanical and physiological meetings pay a considerable amount of attention to plant molecular biology. Since the first edition of this manual appeared, we have published, yearly, a series of supplements to the original volume. These supplements covered new subjects and described new methods that had been developed. With time, however, the editors realized that the original manual plus supplements had become cumbersome to use, and we decided to publish a reorganized version of the manual.

Molecular Biology Techniques: A Classroom Laboratory Manual, Fourth Edition is a must-have collection of methods and procedures on how to create a single, continuous, comprehensive project that teaches students basic molecular techniques. It 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 will gain hands-on experience on subcloning a gene into an expression vector straight through to the purification of the recombinant protein. Presents student-tested labs proven successful in real classroom laboratories Includes a test bank on a companion website for additional testing and practice Provides exercises that simulate a cloning project that would be performed in a real research lab Includes a prep-list appendix that contains necessary recipes and catalog numbers, providing staff with detailed instructions

A comprehensive manual dealing with the molecular aspects of plant molecular biology, which is designed both as a teaching aid for advanced laboratory courses and as a guide for laboratory research. Subjects covered include the introduction of genes into plants, assays of gene activity, plant nucleic acid extraction and characterization procedures, and the use of selectable and screenable markers in transgenic plants. Special emphasis is placed on the use of Agrobacterium tumefaciens to introduce genes into plants, but other gene introduction techniques are also presented. Each chapter, written by experts in the field, includes a discussion of the background to each technique, detailed protocols, media and solution formulas, and an extensive bibliography.