Animal Cell Electroporation and Electrofusion Protocols

Animal Cell Electroporation and Electrofusion Protocols

Author: Jac A. Nickoloff

Publisher: Springer Science & Business Media

Published: 2008-02-03

Total Pages: 367

ISBN-13: 1592595359

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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.


Guide to Electroporation and Electrofusion

Guide to Electroporation and Electrofusion

Author: Donald C. Chang

Publisher: Academic Press

Published: 2012-12-02

Total Pages: 592

ISBN-13: 0080917275

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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


Plant Cell Electroporation And Electrofusion Protocols

Plant Cell Electroporation And Electrofusion Protocols

Author: Jac A. Nickoloff

Publisher: Springer Science & Business Media

Published: 2008-02-02

Total Pages: 209

ISBN-13: 1592595421

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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.


Comparative Animal Biochemistry

Comparative Animal Biochemistry

Author: Klaus Urich

Publisher: Springer Science & Business Media

Published: 1994-09-16

Total Pages: 810

ISBN-13: 9783540574200

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There exists an extreme variability in the biochemical properties of animal organisms. Each species, each individual, in fact each cell type of a multicellular animal has its own molecular characters. This advanced text- and reference book deals with the molecular structures of body substances and the chemical processes of metabolism for the first time throughout the whole animal kingdom from the protozoans to the higher vertebrates. The biochemical variation is explained as adaptation to particular environmental conditions or as the result of phylogenetic diversification. The fascinating insights into the fundamental mechanisms and the time course of evolution that have been gained by the analysis of molecular data are extensively described. More than 4600 actual references give the reader access to the original literature.


Computer Analysis of Sequence Data Part II

Computer Analysis of Sequence Data Part II

Author: Annette M. Griffin

Publisher: Springer Science & Business Media

Published: 2008-02-02

Total Pages: 434

ISBN-13: 159259512X

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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.


Capillary Electrophoresis Guidebook

Capillary Electrophoresis Guidebook

Author: Kevin D. Altria

Publisher: Springer Science & Business Media

Published: 2008-02-02

Total Pages: 347

ISBN-13: 1592595391

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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.


Biological and Medical Aspects of Electromagnetic Fields, Fourth Edition

Biological and Medical Aspects of Electromagnetic Fields, Fourth Edition

Author: Ben Greenebaum

Publisher: CRC Press

Published: 2018-11-01

Total Pages: 651

ISBN-13: 1351737724

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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


Biological and Medical Aspects of Electromagnetic Fields

Biological and Medical Aspects of Electromagnetic Fields

Author: Frank S. Barnes

Publisher: CRC Press

Published: 2018-10-03

Total Pages: 478

ISBN-13: 142000946X

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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.


Handbook of Biochemistry and Molecular Biology

Handbook of Biochemistry and Molecular Biology

Author:

Publisher: CRC Press

Published: 2010-05-21

Total Pages: 1100

ISBN-13: 1466515155

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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.


Transgenic Animals

Transgenic Animals

Author: Louis-Marie Houdebine

Publisher: CRC Press

Published: 2022-04-19

Total Pages: 122

ISBN-13: 1000448436

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During the past 20 years, transgenesis has become a popular technique and a crucial tool for molecular geneticists and biologists. Transgene expression is now better-controlled and even specifically inducible by exogenous factors. While these techniques have quite significantly transformed the experimental approaches taken by biologists, the applications are more limited than expected and concerns have arisen regarding biosafety as well as physiological, social, and philosophical issues. Transgenic Animals: Generation and Use contains articles on the techniques used to generate transgenic animals and a section on the preparation of vectors for the optimally controlled expression of transgenes. It also examines the use of transgenic animals in the study of gene function and human diseases, the preparation of recombinant proteins and organs for pharmaceutical and medical use, and the improvement of genetic characteristics of farm animals. Finally, it discusses more recent problems generated by transgenic animals including conservation of transgenic lines, specific database patenting, biosafety, and bioethics. Drawn from both academia and industry, the contributors to this monograph present in one concise volume all the relevant information on the different aspects of transgenesis. This book can be used as both a reference book and a textbook for specialized university courses and will be of interest to everyone involved in basic research in animal biology, molecular genetics, animal biotechnology, pharmaceutical science, and medicine.