In Vitro Mutagenesis Protocols

Author: Michael K. Trower

Publisher: Springer Science & Business Media


Category: Science

Page: 390

View: 4432

In In Vitro Mutagenesis Protocols leading experts from industrial and academic laboratories describe easily reproducible procedures for site-directed and random mutagenesis. Site-directed protocols include those based on strand-selection, PCR (including "splicing by overlap extension" and the "megaprimer" procedure), the ligase chain reaction, positive antibiotic selection, unique restriction site elimination, gapped heteroduplex formation, and solid-phase capture with the biotin/ strepavidin system. Many techniques can be used with virtually any double-stranded DNA plasmid. The random mutagenesis protocols include methods based on PCR, degenerate oligonucleotides, cassette mutagenesis, nested deletion mutagenesis, and a specialized E. coli mutator strain. These invaluable protocols facilitate the study of gene regulation and structure/function relationships in proteins and permit modification of DNA sequences for purposes such as vector construction.

In Vitro Mutagenesis Protocols

Author: Jeff Braman

Publisher: Springer Science & Business Media

ISBN: 1592591949

Category: Science

Page: 287

View: 3422

In the post-genome era, in vitro mutagenesis has emerged as the critically important tool used by molecular biologists in establishing the functions of components of the proteome. In this second edition of In Vitro Mutagenesis Protocols, active researchers with proven track records describe in stepwise fashion their advanced mutagenesis techniques. Each contributor focuses on improvements to conventional site-directed mutagenesis, with chapters being devoted to chemical site-directed mutagenesis; PCR-based mutagenesis and the modifications that allow high-throughput experiments; and mutagenesis based on gene disruption that is both in vitro- and in situ-based. Additional methods are provided for in vitro gene evolution; for gene disruption based on transposon, recombination, and cassette mutagenesis; and for facilitating the introduction of multiple mutations. Each readily reproducible technique includes detailed step-by-step instructions, tips on pitfalls to avoid, and notes on reagents and suppliers. Time-tested and highly practical, the techniques in In Vitro Mutagenesis Protocols, Second Edition offer today's molecular biologists a rich compendium of reliable and powerful techniques with which to illuminate the proteome.

In Vitro Mutagenesis: Methods and Protocols

Author: Andrew Reeves

Publisher: Methods in Molecular Biology

ISBN: 9781493982110

Category: Medical

Page: 528

View: 7308

In vitro mutagenesis remains a critical experimental approach for investigating gene and protein function at the cellular level. This volume provides a wide variety of updated and novel approaches for performing in vitro mutagenesis using such methods as genome editing, transposon (Tn) mutagenesis, site-directed, and random mutagenesis. In Vitro Mutagenesis: Methods and Protocols guides readers through methods for gene and genome editing, practical bioinformatics approaches for identifying mutagenesis targets, and novel site-directed and random mutagenesis approaches aimed at gaining a better understanding of protein-protein and protein-cofactor interactions. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, In Vitro Mutagenesis: Methods and Protocols aims to provide a highly accessible and practical manual for current and future molecular biology researchers, from the beginner practitioner to the advanced investigator in fields such as molecular genetics, biochemistry, and biochemical and metabolic engineering.


Analysis and Design

Author: Ruth Hogue Angeletti

Publisher: Elsevier

ISBN: 0080539998

Category: Science

Page: 362

View: 2144

Proteins: Analysis and Design focuses solely on individual experimental approaches, rather than on specific classes of proteins. The book provides insight into the important issues in protein science and how one can cope with them. These include all issues which explore the detailed relationship of protein structure to function. Provides problems and technical solutions Includes posttranslational modifications Uses synthetic peptides as biological models Details mutagenesis and protein engineering Covers design of protein structure and function

Comprehensive Biomaterials II

Author: Paul Ducheyne,Kevin Healy,Dietmar W. Hutmacher,David W. Grainger,C. James Kirkpatrick

Publisher: Elsevier

ISBN: 0081006926

Category: Technology & Engineering

Page: 4858

View: 1828

Comprehensive Biomaterials II, Second Edition brings together the myriad facets of biomaterials into one expertly-written series of edited volumes. Articles address the current status of nearly all biomaterials in the field, their strengths and weaknesses, their future prospects, appropriate analytical methods and testing, device applications and performance, emerging candidate materials as competitors and disruptive technologies, research and development, regulatory management, commercial aspects, and applications, including medical applications. Detailed coverage is given to both new and emerging areas and the latest research in more traditional areas of the field. Particular attention is given to those areas in which major recent developments have taken place. This new edition, with 75% new or updated articles, will provide biomedical scientists in industry, government, academia, and research organizations with an accurate perspective on the field in a manner that is both accessible and thorough. Reviews the current status of nearly all biomaterials in the field by analyzing their strengths and weaknesses, performance, and future prospects Covers all significant emerging technologies in areas such as 3D printing of tissues, organs and scaffolds, cell encapsulation; multimodal delivery, cancer/vaccine - biomaterial applications, neural interface understanding, materials used for in situ imaging, and infection prevention and treatment Effectively describes the many modern aspects of biomaterials from basic science, to clinical applications

Microbial Biotechnology

Principles and Applications Third

Author: Yuan Kun Lee

Publisher: World Scientific Publishing Company

ISBN: 9814513091

Category: Science

Page: 896

View: 7888

The rapidly expanding molecular biological techniques and approaches have significant impact on microbial biotechnology, hence the need for the addition of four new chapters in the third edition of this textbook — “Chapter 3: Application of ‘Omics’ Technologies in Microbial Fermentation”, “Chapter 5: Microbial Genome Mining for Identifying Antimicrobial Targets”, “Chapter 21: Bacterial Biofilm: Molecular Characterization and Impacts on Water Management” and “Chapter 23: Microbial Biomining”. “Chapter 15: Transgenic Plants” has been completely revised while most of the other chapters have been thoroughly updated in this new edition. There already exist a number of excellent general textbooks on microbiology and biotechnology that deal with the basic principles of microbial biotechnology. To complement them, this book focuses on the various applications of microbial-biotechnological principles. A teaching-based format is adopted, whereby working problems, as well as answers to frequently asked questions, supplement the main text. The book also includes real life examples of how the application of microbial-biotechnological principles has achieved breakthroughs in both research and industrial production. Although written for polytechnic students and undergraduates, the book contains sufficient information to be used as a reference for postgraduate students and lecturers. It may also serve as a resource book for corporate planners, managers and applied research personnel.

Yeast Protocols

Author: Wei Xiao

Publisher: Springer Science & Business Media

ISBN: 1592599583

Category: Science

Page: 392

View: 6995

Since the publication of Yeast Protocols in 1996, many new techniques have been invented and original protocols improved and refined. This throughly updated second edition will serve as a stand-alone protocols handbook of these new and refined techniques suitable for daily use in all research laboratories. It includes all of the recent advanced protocols as well as the major basic techniques and hence, will be essential for both yeast research laboratories and those researchers who wish to use yeast as a host to study their favorite genes from other organisms.

Cell Cycle Checkpoint Control Protocols

Author: Howard B. Lieberman

Publisher: Springer Science & Business Media

ISBN: 1592596460

Category: Science

Page: 372

View: 952

A collection of basic cutting-edge techniques for studying the mechanisms underlying cell cycle regulation and checkpoint control. Using mammalian, yeast, and frog systems, these readily reproducible methods can be used to induce cell cycle checkpoints, detect changes in cell cycle progression, identify and analyze genes and proteins that regulate the process, and characterize chromosomal status as a function of cell cycle phase and progression. Each fully tested technique includes step-by-step instructions written by an investigator who routinely performs it, an introduction explaining the principle behind the method, equipment and reagent lists, and tips on troubleshooting and avoiding known pitfalls.

Comprehensive Biomaterials

Author: Paul Ducheyne,Kevin Healy,Dietmar W. Hutmacher,David W. Grainger,C. James Kirkpatrick

Publisher: Elsevier

ISBN: 0080552943

Category: Technology & Engineering

Page: 3672

View: 6665

Comprehensive Biomaterials brings together the myriad facets of biomaterials into one, major series of six edited volumes that would cover the field of biomaterials in a major, extensive fashion: Volume 1: Metallic, Ceramic and Polymeric Biomaterials Volume 2: Biologically Inspired and Biomolecular Materials Volume 3: Methods of Analysis Volume 4: Biocompatibility, Surface Engineering, and Delivery Of Drugs, Genes and Other Molecules Volume 5: Tissue and Organ Engineering Volume 6: Biomaterials and Clinical Use Experts from around the world in hundreds of related biomaterials areas have contributed to this publication, resulting in a continuum of rich information appropriate for many audiences. The work addresses the current status of nearly all biomaterials in the field, their strengths and weaknesses, their future prospects, appropriate analytical methods and testing, device applications and performance, emerging candidate materials as competitors and disruptive technologies, and strategic insights for those entering and operational in diverse biomaterials applications, research and development, regulatory management, and commercial aspects. From the outset, the goal was to review materials in the context of medical devices and tissue properties, biocompatibility and surface analysis, tissue engineering and controlled release. It was also the intent both, to focus on material properties from the perspectives of therapeutic and diagnostic use, and to address questions relevant to state-of-the-art research endeavors. Reviews the current status of nearly all biomaterials in the field by analyzing their strengths and weaknesses, performance as well as future prospects Presents appropriate analytical methods and testing procedures in addition to potential device applications Provides strategic insights for those working on diverse application areas such as R&D, regulatory management, and commercial development

PCR Protocols

Current Methods and Applications

Author: Bruce Alan White

Publisher: Springer Science & Business Media

ISBN: 1592595022

Category: Cytology

Page: 392

View: 4541

The Nucleic Acid Protocols Handbook

Author: Ralph Rapley

Publisher: Springer Science & Business Media

ISBN: 1592590381

Category: Science

Page: 478

View: 8405

A comprehensive treasury of all the key molecular biology methods-ranging from DNA extraction to gene localization in situ-needed to function effectively in the modern laboratory. Each of the 120 highly successful techniques follows the format of the much acclaimed Methods in Molecular BiologyOao series, providing an introduction to the scientific basis of each technique, a complete listing of all the necessary materials and reagents, and clear step-by-step instruction to permit error-free execution. Included for each technique are notes about pitfalls to avoid, troubleshooting tips, alternate methods, and explanations of the reasons for certain steps-all key elements contributing significantly to success or failure in the lab. The Nucleic Acid Protocols Handbook constitutes today's most comprehensive collection of all the key classic and cutting-edge techniques for the successful isolation, analysis, and manipulation of nucleic acids by both experienced researchers and those new to the field."

Molecular Toxicology Protocols

Author: Phouthone Keohavong,Stephen G. Grant

Publisher: Springer Science & Business Media

ISBN: 1592598404

Category: Medical

Page: 489

View: 7669

New state-of-the-art molecular techniques promise to transform the field of genetic toxicology by making it possible to directly detect genotoxic exposures and their consequences in humans, to identify the agent(s) involved, and to clinically manage the exposed population. In Molecular Toxicology Protocols, researchers from prominent universities and cancer centers around the world describe in detail their best techniques for analyzing genotoxic exposure and the resulting biological effects, including intermediate biomarkers such as DNA and chromosomal damage, mutations in reporter and oncogenes, and the earliest possible detection of cancer. The authors emphasize analytical methods specifically developed for use in human populations and in cancer patients, or in other in vivo systems such as transgenic mice. Among the applications detailed are the analysis of interactions of chemical and physical agents with cellular macromolecules, especially DNA, the assessment of medically relevant toxicity, and the individualized characterization of genetic damage and repair. There are also methods to assess and characterize the modulation of this damage and repair through individual differences in specific and genome-wide gene expression, including metabolic profiling and apoptotic capacity. These methods mark a shift in emphasis from studies of the agents themselves to the exposed population, and from studies of small populations with significant known exposures to a single agent, to studies of common diseases, such as breast cancer, caused by normal levels of generalized genotoxic exposure. The protocols follow the successful Methods in Molecular Biology series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and highly practical, Molecular Toxicology Protocols offers a gold-standard collection of cutting-edge techniques designed to investigate a broad range of exposures-endogenous, accidental, medical, environmental, and occupational-and their role in human carcinogenesis and other diseases of aging.

PCR Cloning Protocols

Author: Bing-Yuan Chen,Harry W. Janes

Publisher: Springer Science & Business Media

ISBN: 1592591779

Category: Science

Page: 460

View: 2837

Annotation PCR Cloning Protocols, Second Edition, updates and expands Bruce White's best-selling PCR Cloning Protocols (1997) with the newest procedures for DNA cloning and mutagenesis. Here the researcher will find readily reproducible methods for all the major aspects of PCR use, including PCR optimization, computer programs for PCR primer design and analysis, and novel variations for cloning genes of special characteristics or origin, with emphasis on long distance PCR and GC-rich template amplification. Also included are both conventional and novel enzyme-free and restriction site-free procedures to clone PCR products into a range of vectors, as well as state-of-the-art protocols to facilitate DNA mutagenesis and recombination, and to clone the challenging uncharacterized DNA flanking a known DNA fragment.

Epithelial Cell Culture Protocols

Author: Clare Wise

Publisher: Springer Science & Business Media

ISBN: 9780896038936

Category: Medical

Page: 405

View: 3405

Well-versed experimenters and clinical researchers share their best methods for establishing and maintaining epithelial cell cultures, for analyzing and studying their characteristics, and for using them to set up models of critical biological systems. The emphasis is on the analysis and assessment of epithelial cells, for example by looking at apoptosis and integrins, or by measuring membrane capacitance and confluence. Also described in step-by-step detail are co-culture techniques valuable in developing models for investigating many different in vitro systems, including the blood-brain barrier, drug uptake, and the interaction of epithelial cells with bacteria. Epithelial Cell Culture Protocols offers a step-by-step guide toward a deeper understanding of cellular and molecular mechanisms, as well as a set of robust techniques for specifically evaluating the nature and behavior of epithelial cells.

Biology and Breeding of Food Legumes

Author: Aditya Pratap,Jitendra Kumar

Publisher: CABI

ISBN: 184593766X

Category: Technology & Engineering

Page: 418

View: 6854

Food legumes are important constituents of the human diet and animal feed where they are crucial to a balanced diet, supplying high quality proteins. These crops also play an important role in low-input agricultural production systems by fixing atmospheric nitrogen. Despite systematic and continuous breeding efforts through conventional methods, substantial genetic gains have not been achieved. With the rise in demand for food legumes/pulses and increased market value of these crops, research has focused on increasing production and improving the quality of pulses for both edible and industrial purposes. "Biology and Breeding of Food Legumes" covers the history, origin and evolution, botany, breeding objectives and procedures, nutritional improvement, industrial uses and post-harvest technology and also recent developments made through biotechnological intervention.

The Condensed Protocols from Molecular Cloning : a Laboratory Manual

Author: Joseph Sambrook,David William Russell

Publisher: CSHL Press

ISBN: 0879697717

Category: Science

Page: 800

View: 3941

The Condensed Protocols From Molecular Cloning: A Laboratory Manualis a single–volume adaptation of the three–volume third edition of Molecular Cloning: A Laboratory Manual.This condensed book contains only the step–by–step portions of the protocols, accompanied by selected appendices from the world's best–selling manual of molecular biology techniques. Each protocol is cross–referenced to the appropriate pages in the original manual. This affordable companion volume, designed for bench use, offers individual investigators the opportunity to have their own personal collection of short protocols from the essential Molecular Cloning.

Plant Cell Culture Protocols

Author: Robert D. Hall

Publisher: Springer Science & Business Media

ISBN: 1592595839

Category: Science

Page: 421

View: 8513

In Plant Cell Culture Protocols, Robert Hall and a panel of expert researchers present a comprehensive collection of the most frequently used and broadly applicable techniques for plant cell and tissue culture. Readily reproducible and extensively annotated, the methods cover culture initiation, maintenance, manipulation, application, and long-term storage, with emphasis on techniques for genetic modification and micropropagation. Many of these protocols are currently used in major projects designed to produce improved varieties of important crop plants. In addition, a number of specialized protocols have been included to illustrate the diversity of the techniques available and their widespread applicability. Plant Cell Culture Protocols is aimed at scientists involved in all aspects of plant biotechnological research, as well as those working in other areas of agriculture and horticulture who are interested in expanding their technical repertoire to include in vitro methodology. Its state-of-the-art techniques are certain to make the book today's reference of choice, an indispensable tool in the development of new transgenic plants and full-scale commercial applications.

Gene Isolation and Mapping Protocols

Author: Jacqueline Boultwood

Publisher: Humana PressInc


Category: Science

Page: 318

View: 1756

An unprecedented collection of all the most up-to-date techniques for gene isolation and mapping, including the latest methods for gene characterization using database analyses. This collection of thoroughly tested recipes also includes chapters for the computational analysis of novel cDNA sequences with up-to-the-minute information on basic sequence analysis, sequence similarity searches, exon detection and similarity searches, and the prediction of gene function. Its state-of-the-art methods constitute indispensable tools for all scientists engaged in the search for specific disease genes, or in the general advancement of the human genome project.