Animal Cell Electroporation and Electrofusion Protocols PDF Download
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Author: Jac A. Nickoloff
Publisher: Springer Science & Business Media
ISBN: 1592595359
Category : Science
Languages : en
Pages : 367
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Book Description
The ability to introduce macromolecules into animal cells, includ ing DNA, RNA, proteins, and other bioactive compounds has facili tated a broad range of biological studies, from biochemistry and biophysics to molecular biology, cell biology, and whole animal stud ies. Gene transfer technology in particular will continue to play an essential role in studies aimed at improving our understanding of the relationships between the gene structure and function, and it has impor tant practical applications in both biotechnology and biomedicine, as evidenced by the current intense interest in gene therapy. Although DNA and other macromolecules may be introduced into cells by a variety of methods, including chemical treatments and microinjection, el- troporation has proven to be simpler to perform, more efficient, and effective with a wider variety of cell types than other techniques. The early and broad success of electric field-mediated DNA transfer soon prompted researchers to investigate electroporation for transferring other types of molecules into cells, including RNA, enzymes, antibodies, and analytic dyes. Animal Cell Electroporation and Electrofusion Protocols begins with three chapters that describe the theoretical and practical aspects of electroporation, including a review of the commercially available instrumentation. These introductory chapters will be of particular inter est to those new to electric field technologies and to those developing protocols for as yet untested species or cell types. Nineteen chapters follow that present well-tested protocols for electroporation of proteins and DNA into insect, fish, and mammalian cells.
Author: Jac A. Nickoloff
Publisher: Springer Science & Business Media
ISBN: 1592595359
Category : Science
Languages : en
Pages : 367
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Book Description
The ability to introduce macromolecules into animal cells, includ ing DNA, RNA, proteins, and other bioactive compounds has facili tated a broad range of biological studies, from biochemistry and biophysics to molecular biology, cell biology, and whole animal stud ies. Gene transfer technology in particular will continue to play an essential role in studies aimed at improving our understanding of the relationships between the gene structure and function, and it has impor tant practical applications in both biotechnology and biomedicine, as evidenced by the current intense interest in gene therapy. Although DNA and other macromolecules may be introduced into cells by a variety of methods, including chemical treatments and microinjection, el- troporation has proven to be simpler to perform, more efficient, and effective with a wider variety of cell types than other techniques. The early and broad success of electric field-mediated DNA transfer soon prompted researchers to investigate electroporation for transferring other types of molecules into cells, including RNA, enzymes, antibodies, and analytic dyes. Animal Cell Electroporation and Electrofusion Protocols begins with three chapters that describe the theoretical and practical aspects of electroporation, including a review of the commercially available instrumentation. These introductory chapters will be of particular inter est to those new to electric field technologies and to those developing protocols for as yet untested species or cell types. Nineteen chapters follow that present well-tested protocols for electroporation of proteins and DNA into insect, fish, and mammalian cells.
Author: Donald C. Chang
Publisher: Academic Press
ISBN: 0080917275
Category : Science
Languages : en
Pages : 592
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Book Description
Electroporation is an efficient method to introduce macromolecules such as DNA into a wide variety of cells. Electrofusion results in the fusion of cells and can be used to produce genetic hybrids or hybridoma cells.Guide to Electroporation and Electrofusion is designed to serve the needs of students, experienced researchers, and newcomers to the field. It is a comprehensive manual that presents, in one source, up-to-date, easy-to-follow protocols necessary for efficient electroporation and electrofusion of bacteria, yeast, and plant and animal cells, as well as background information to help users optimize their results through comprehension of the principles behind these techniques. - Covers fundamentals of electroporation and electrofusion in detail: Molecular events, Mechanisms, Kinetics, Gives extensive practical information, The latest applications, Controlling parameters to maximize efficiency, Available instrumentation - Presents applications of electroporation and electrofusion in current research situations - State-of-the-art modifications to electrical pulses and generators - Application of electroporation and electrofusion to unique, alternative cell and tissue types - Gives straightforward, detailed, easy-to-follow protocols for Formation of human hybridomas - Introduction of genetic material into plant cells and pollen - Transfection of mammalian cells - Transformation of bacteria, plants, and yeast - Production of altered embryos - Optimization of electroporation by using reporter genes - Comprehensive and up-to-date - Convenient bench-top format - Approximately 125 illustrations complement the text - Complete references with article titles - Written by leading authorities in electroporation and electrofusion
Author: Jac A. Nickoloff
Publisher: Springer Science & Business Media
ISBN: 1592595421
Category : Science
Languages : en
Pages : 209
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Book Description
Gene transfer is an essential technology for improving our under standing of gene structure and function. Although there are many meth ods by which DNA may be introduced into cells—including heat and chemical treatments, and microinjection—electroporation has been found to be the most versatile gene transfer technique. Electroporation is effective with a wide variety of cell types, including those that are difficult to transform by other means. For many cell types, electroporation is either the most efficient or the only means known to effect gene transfer. The early and broad success of electric field-medi ated DNA transfer soon prompted researchers to investigate electroporation for transferring other types of molecules into cells, in cluding RNA, enzymes, antibodies, and analytic dyes. The first section of Plant Cell Electroporation and Electrofusion Protocols includes two chapters that serve as a guide to theoretical and practical aspects of electroporation, and will be of particular interest to those developing protocols for as yet untested species or cell types, and a third chapter that describes commercially available electroporation instruments. The remaining chapters describe well-tested protocols for DNA electrotransfection, electroporation of other biomolecules, or cell electrofusion. These chapters also include brief discussions of alterna tives to electric field-based methods, citing the advantages and limita tions of the various methods for achieving specific goals.
Author: Annette M. Griffin
Publisher: Springer Science & Business Media
ISBN: 159259512X
Category : Science
Languages : en
Pages : 434
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Book Description
DNA sequencing has become increasingly efficient over the years, resulting in an enormous increase in the amount of data gener ated. In recent years, the focus of sequencing has shifted, from being the endpoint of a project, to being a starting point. This is especially true for such major initiatives as the human genome project, where vast tracts of DNA of unknown function are sequenced. This sheer volume of available data makes advanced computer methods essen tial to analysis, and a familiarity with computers and sequence analy sis software a vital requirement for the researcher involved with DNA sequencing. Even for nonsequencers, a familiarity with sequence analysis software can be important. For instance, gene sequences already present in the databases can be extremely useful in the design of cloning and genetic manipulation experiments. This two-part work on Computer Analysis of Sequence Data is designed to be a practical aid to the researcher who uses computers for the acquisition, storage, or analysis of nucleic acid (and/or pro tein) sequences. Each chapter is written such that a competent scien tist with basic computer literacy can carry out the procedure successfully at the first attempt by simply following the detailed prac tical instructions that have been described by the author. A Notes section, which is included at the end of each chapter, provides advice on overcoming the common problems and pitfalls sometimes encoun tered by users of the sequence analysis software.
Author: Kevin D. Altria
Publisher: Springer Science & Business Media
ISBN: 1592595391
Category : Science
Languages : en
Pages : 347
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Book Description
This book is intended to be a working guide to the operation of capillary electrophoresis (CE) instrumentation. Since CE is still a rap idly maturing technique, detailed validated protocols are not widely established. Therefore, extensive experimental procedures are not pro vided for individual analyses. The intention is to provide general guide lines on the principles and practice of CE and to give an overview of the specific technologies and important application areas. Part I provides operating instructions for standard commercially available instruments. Guidelines are included for activities such as changing capillaries, method development, quantitative procedures, optimization of precision and sensitivity, and the validation of meth ods, fraction collection, and troubleshooting, as well as a quick guide to running a separation. The application range of CE is possibly the most diverse of all analytical techniques and ranges from large, complex macromolecules, such as proteins and nucleic acids, to small solutes, such as organic drugs and inorganic anions and cations.
Author: Ben Greenebaum
Publisher: CRC Press
ISBN: 1351737724
Category : Technology & Engineering
Languages : en
Pages : 651
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Book Description
The two volumes of this new edition of the Handbook cover the basic biological, medical, physical, and electrical engineering principles. They also include experimental results concerning how electric and magnetic fields affect biological systems—both as potential hazards to health and potential tools for medical treatment and scientific research. They also include material on the relationship between the science and the regulatory processes concerning human exposure to the fields. Like its predecessors, this edition is intended to be useful as a reference book but also for introducing the reader to bioelectromagnetics or some of its aspects. FEATURES • New topics include coverage of electromagnetic effects in the terahertz region, effects on plants, and explicitly applying feedback concepts to the analysis of biological electromagnetic effects • Expanded coverage of electromagnetic brain stimulation, characterization and modeling of epithelial wounds, and recent lab experiments on at all frequencies • Section on background for setting standards and precautionary principle • Discussion of recent epidemiological, laboratory, and theoretical results; including: WHO IARC syntheses of epidemiological results on both high and low frequency fields, IITRI lab study of cancer in mice exposed to cell phone-like radiation, and other RF studies • All chapters updated by internationally acknowledged experts in the field
Author: Frank S. Barnes
Publisher: CRC Press
ISBN: 142000946X
Category : Technology & Engineering
Languages : en
Pages : 478
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Book Description
Biological and Medical Aspects of Electromagnetic Fields examines potential health hazards, exposure standards, and medical applications of electromagnetic (EM) fields. The second volume in the bestselling and newly revisedHandbook of Biological Effects of Electromagnetic Fields, Third Edition, this book draws from the latest studies on the effects of exposure to electric and magnetic fields. In addition to extensive reviews of physiological effects, the book contains now separate reviews of behavioral and cognitive responses to various exposures. The book also describes an approach to setting standards for exposure limits and explores a few of the beneficial uses of EM fields in medical applications, both diagnostics and in treatment. Biological and Medical Aspects of Electromagnetic Fields provides a practical overview of the experiments and methods used to observe ELF and RF fields and the possible useful and hazardous implications of these observations.
Author:
Publisher: CRC Press
ISBN: 1466515155
Category : Medical
Languages : en
Pages : 1100
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Book Description
Edited by renowned protein scientist and bestselling author Roger L. Lundblad, with the assistance of Fiona M. Macdonald of CRC Press, this fourth edition of the Handbook of Biochemistry and Molecular Biology represents a dramatic revision — the first in two decades — of one of biochemistry's most referenced works. This edition gathers a wealth of information not easily obtained, including information not found on the web. Offering a molecular perspective not available 20 years ago, it provides physical and chemical data on proteins, nucleic acids, lipids, and carbohydrates. Presented in an organized, concise, and simple-to-use format, this popular reference allows quick access to the most frequently used data. Covering a wide range of topics, from classical biochemistry to proteomics and genomics, it also details the properties of commonly used biochemicals, laboratory solvents, and reagents. Just a small sampling of the wealth of information found inside the handbook: Buffers and buffer solutions Heat capacities and combustion levels Reagents for the chemical modification of proteins Comprehensive classification system for lipids Biological characteristics of vitamins A huge variety of UV data Recommendations for nomenclature and tables in biochemical thermodynamics Guidelines for NMR measurements for determination of high and low pKa values Viscosity and density tables Chemical and physical properties of various commercial plastics Generic source-based nomenclature for polymers Therapeutic enzymes About the Editors: Roger L. Lundblad, Ph.D. Roger L. Lundblad is a native of San Francisco, California. He received his undergraduate education at Pacific Lutheran University and his PhD degree in biochemistry at the University of Washington. After postdoctoral work in the laboratories of Stanford Moore and William Stein at the Rockefeller University, he joined the faculty of the University of North Carolina at Chapel Hill. He joined the Hyland Division of Baxter Healthcare in 1990. Currently Dr. Lundblad is an independent consultant and writer in biotechnology in Chapel Hill, North Carolina. He is an adjunct Professor of Pathology at the University of North Carolina at Chapel Hill and Editor-in-Chief of the Internet Journal of Genomics and Proteomics. Fiona M. Macdonald, Ph.D., F.R.S.C. Fiona M. Macdonald received her BSc in chemistry from Durham University, UK. She obtained her PhD in inorganic biochemistry at Birkbeck College, University of London, studying under Peter Sadler. Having spent most of her career in scientific publishing, she is now at Taylor and Francis and is involved in developing chemical information products.
Author: Klaus Urich
Publisher: Springer Science & Business Media
ISBN: 3662063034
Category : Science
Languages : en
Pages : 798
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Book Description
tribute greatly to understanding the origins of The plan for this book goes back almost 20 years. Already, at that time, it was possible to recognize organisms. an extraordinary variation in metabolites and To provide the biochemist with a ready over processes superimposed upon the basic biochem view of the structural diversity of animals, the book includes a simplified version of animal sys ical system of animals. Each species, each indi tematics; for further information on the classifica vidual, in fact each type of cell of the multicellu lar organism possesses its own biochemical char tion, structure and life of particular animal spe acter, and this molecular variety, its biological sig cies, the reader should consult the relevant text nificance, and its evolutionary development books. It is assumed that the zoologist reader has throw up many interesting questions. The com a basic knowledge of biochemistry; important general biochemical facts are in any case given for parative approach that has been so productive at many of the subjects covered. the higher levels of complexity of morphology and physiology can also be used to great effect at I had already completed several chapters of the molecular level. this book by the beginning of the 1970s.
Author: M. A Titheradge
Publisher: Springer Science & Business Media
ISBN: 1592597491
Category : Science
Languages : en
Pages : 321
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Book Description
Michael Titheradge provides both the experienced researcher and the newcomer with time-tested techniques to cover all aspects of NO synthase and metabolism. Topics range from the cloning and expression of the different isoforms of NO synthase to the role of NO in DNA damage and apoptosis and include indirect and direct measurements of NO production, measurement on NO synthase mRNA both in situ and in vitro, and Western blotting and immunohistochemical localization of different isoforms of NO synthase. A variety of alternative techniques are described to enable researchers to set up an assay using either the simplest equipment or the most expensive EPR machines.
