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This book offers step-by-step instruction on DNA cloning, defined as moving genes around plasmids, mutating genes, or mining new genes. The aim is to provide those new to the field with reliable and up-to-date practical guidance while at the same time conveying the scope for creativity. After a brief synopsis of the history of cloning, the fundamentals and prerequisites are explained, covering, for example, software, vectors commonly used in the lab, appropriate choice of restriction endonucleases, the preparation of agarose gels, competent cells, and LB agar plates, and procedures to be followed upon receipt of new plasmids. The remainder of the book is devoted to the clear description of methods and individual steps in cloning. Guidance is provided on the cut and paste method, DNA sequencing, direct sequencing, primer design, PCR-based gene insertion and deletion, epitope tag insertion, the use of RACE technology, BAC recombineering, and much, much more. Sources of error and a variety of techniques that make life considerably easier when cloning are also examined in detail.
Human reproductive cloning is an assisted reproductive technology that would be carried out with the goal of creating a newborn genetically identical to another human being. It is currently the subject of much debate around the world, involving a variety of ethical, religious, societal, scientific, and medical issues. Scientific and Medical Aspects of Human Reproductive Cloning considers the scientific and medical sides of this issue, plus ethical issues that pertain to human-subjects research. Based on experience with reproductive cloning in animals, the report concludes that human reproductive cloning would be dangerous for the woman, fetus, and newborn, and is likely to fail. The study panel did not address the issue of whether human reproductive cloning, even if it were found to be medically safe, would beâ€"or would not beâ€"acceptable to individuals or society.
In DNA Cloning and Assembly Methods, expert researchers in the field detail many of the methods which are now commonly used for DNA cloning and make cloning procedures faster, more reliable and also suitable for high-throughput handling. These include methods and protocols that are based on several mechanisms including type II and IIS restriction enzymes, single stranded annealing, sequence overlap, and recombination. With additional chapters on software programs that are suitable for primer design, a feature crucial for the functionality of the described methods. 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 key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, DNA Cloning and Assembly Methods seeks to provide scientist with a valuable and useful resource for wet lab researchers within life sciences.
Known world-wide as the standard introductory text to this important and exciting area, the sixth edition of Gene Cloning and DNA Analysis addresses new and growing areas of research whilst retaining the philosophy of the previous editions. Assuming the reader has little prior knowledge of the subject, its importance, the principles of the techniques used and their applications are all carefully laid out, with over 250 clearly presented four-colour illustrations. In addition to a number of informative changes to the text throughout the book, the final four chapters have been significantly updated and extended to reflect the striking advances made in recent years in the applications of gene cloning and DNA analysis in biotechnology. Gene Cloning and DNA Analysis remains an essential introductory text to a wide range of biological sciences students; including genetics and genomics, molecular biology, biochemistry, immunology and applied biology. It is also a perfect introductory text for any professional needing to learn the basics of the subject. All libraries in universities where medical, life and biological sciences are studied and taught should have copies available on their shelves. "... the book content is elegantly illustrated and well organized in clear-cut chapters and subsections... there is a Further Reading section after each chapter that contains several key references... What is extremely useful, almost every reference is furnished with the short but distinct author's remark." –Journal of Heredity, 2007 (on the previous edition)
This volume provides a comprehensive collection of DNA assembly protocols that will prove useful for any researcher interested in molecular cloning, synthetic biology, or DNA manipulation. Chapters will guide readers through computational tools to design and track the construction of DNA assemblies, workflows that enable high-throughput assembly of DNA constructs, standardized toolkits and protocols for DNA assembly, and combinatorial solutions that enable the construction and optimization of entire metabolic pathways. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, application details for both the expert and non-expert reader, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, DNA Cloning and Assembly: Methods and Protocols aims to ensure successful results in the further study of this vital field.
Clear and concise, this easy-to-use text offers an introductory course on the language of gene cloning, covering microbial, plant, and animal systems. The essential concepts in biology relevant to the understanding of gene cloning are presented in a well-organized and accessible manner. This updated version of the first edition is an invaluable book for nonscientists as well as scientists with little background knowledge in gene cloning, providing a wealth of information for anyone wishing to gain proficiency in reading and speaking the language of gene cloning.
Updated to reflect advances in the field, this introduction provides a broad, but concise, coverage of recombinant DNA techniques. Written for advanced undergraduates, graduates and scientists who want to use this technology, emphasis is placed on the concepts underlying particular types of cloning vectors to aid understanding and to enable readers to devise suitable strategies for novel experimental situations. An introduction to the basic biochemical principles is presented first. Then PCR and cloning using E. coli hosts and plasmid, phage and hybrid vectors are described, followed by the generation and screening of libraries and how to modify, inactivate or express cloned sequences. Finally genetic manipulation in a range of other organisms is discussed, including other bacteria, fungi, algae and plants, insects and mammals. A series of 'real-life' biological problems are also presented to enable readers to assess their understanding of the material and to prepare for exams.
An insider's view on bringing extinct species back to life Could extinct species, like mammoths and passenger pigeons, be brought back to life? In How to Clone a Mammoth, Beth Shapiro, an evolutionary biologist and pioneer in ancient DNA research, addresses this intriguing question by walking readers through the astonishing and controversial process of de-extinction. From deciding which species should be restored to anticipating how revived populations might be overseen in the wild, Shapiro vividly explores the extraordinary cutting-edge science that is being used to resurrect the past. Considering de-extinction's practical benefits and ethical challenges, Shapiro argues that the overarching goal should be the revitalization and stabilization of contemporary ecosystems. Looking at the very real and compelling science behind an idea once seen as science fiction, How to Clone a Mammoth demonstrates how de-extinction will redefine conservation's future.
Recombinant DNA methods are powerful, revolutionary techniques that allow the isolation of single genes in large amounts from a pool of thousands or millions of genes and the modification of these isolated genes or their regulatory regions for reintroduction into cells for expression at the RNA or protein levels. These attributes lead to the solution of complex biological problems and the production of new and better products in the areas of medicine, agriculture, and industry. Recombinant DNA Methodology, a volume in the Selected Methods in Enzymology series produced in benchtop format, contains a selection of key articles from Volumes 68, 100, 101, 153, 154, and 155 of Methods in Enzymology. The essential and widely used procedures provided at an affordable price will be an invaluable aid to the graduate student and the researcher. - Enzymes in DNA research - DNA isolation, hybridization, and cloning - DNA sequence analysis - cDNA cloning - Gene products - Identification of cloned genes and mapping of genes - Monitoring cloned gene expression - Cloning and transferring of genes into yeast cells - Cloning and transferring of genes into plant cells - Cloning and transferring of genes into animal cells - Site-directed mutagenesis - Protein engineering - Expression vectors