Electroporation Protocols

Electroporation Protocols PDF Author: Shulin Li
Publisher: Humana Press
ISBN: 9781461496311
Category : Medical
Languages : en
Pages : 0

Get Book Here

Book Description
Electroporation gene therapy, or gene electrotransfer, has evolved greatly over the last few decades as a result of the remarkable progress in genetic sequencing, gene array analysis, gene cloning, gene expression detection, DNA manufacture and discovery and synthesis of siRNA. Electroporation Protocols: Preclinical and Clinical Gene Medicine, Second Edition provides in-depth knowledge on the delivery of naked DNA and small-interfering RNA (siRNA) to the targeted cells, tissues, and animals for prevention and treatment of disease. It builds on the success of the first edition and on the progress made in siRNA delivery and DNA vaccines for large animals as well as discovery of electroporation applications for the fragile tissues and for internal organs. Written in the 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 protocols and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Electroporation Protocols: Preclinical and Clinical Gene Medicine, Second Edition aims to provide not only comprehensive coverage of the basic theory and practical application of electroporation siRNA therapy, gene therapy, and vaccine, but also elaborates on the most current views from the experts in this field, serving as an invaluable resource for investigators both in and outside of this field.

Electroporation Protocols

Electroporation Protocols PDF Author: Shulin Li
Publisher: Humana Press
ISBN: 9781461496311
Category : Medical
Languages : en
Pages : 0

Get Book Here

Book Description
Electroporation gene therapy, or gene electrotransfer, has evolved greatly over the last few decades as a result of the remarkable progress in genetic sequencing, gene array analysis, gene cloning, gene expression detection, DNA manufacture and discovery and synthesis of siRNA. Electroporation Protocols: Preclinical and Clinical Gene Medicine, Second Edition provides in-depth knowledge on the delivery of naked DNA and small-interfering RNA (siRNA) to the targeted cells, tissues, and animals for prevention and treatment of disease. It builds on the success of the first edition and on the progress made in siRNA delivery and DNA vaccines for large animals as well as discovery of electroporation applications for the fragile tissues and for internal organs. Written in the 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 protocols and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Electroporation Protocols: Preclinical and Clinical Gene Medicine, Second Edition aims to provide not only comprehensive coverage of the basic theory and practical application of electroporation siRNA therapy, gene therapy, and vaccine, but also elaborates on the most current views from the experts in this field, serving as an invaluable resource for investigators both in and outside of this field.

Electroporation Protocols for Microorganisms

Electroporation Protocols for Microorganisms PDF Author: Jac A. Nickoloff
Publisher: Springer Science & Business Media
ISBN: 1592595340
Category : Science
Languages : en
Pages : 374

Get Book Here

Book Description
Electroporation is one of the most widespread techniques used in modem molecular genetics. It is most commonly used to introduce DNA into cells for investigations of gene structure and function, and in this regard, electroporation is both highly versatile, being effective with nearly all species and cell types, and highly efficient. For many cell types, electroporation is either the most efficient or the only means known to effect gene transfer. However, exposure of cells to brief, hi- intensity electric fields has found broad application in other aspects of biological research, and is now routinely used to introduce other types of biological and analytic molecules into cells, to induce cell-cell fusion, and to transfer DNA directly between different species. The first seven chapters of Electroporation Protocols for Micro organisms describe the underlying theory of electroporation, the com mercially available instrumentation, and a number of specialized electroporation applications, such as cDNA library construction and interspecies DNA electrotransfer. Each of the remaining chapters pre sents a well developed method for electrotransformation of a particular bacterial, fungal, or protist species. These chapters also serve to intro duce those new to the field the important research questions that are currently being addressed with particular organisms, highlighting both the major advantages and limitations of each species as a model organ ism, and explaining the roles that electroporation has played in the development of the molecular genetic systems currently in use.

Guide to Electroporation and Electrofusion

Guide to Electroporation and Electrofusion PDF Author: Donald C. Chang
Publisher: Academic Press
ISBN: 0080917275
Category : Science
Languages : en
Pages : 592

Get Book Here

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

Transcription Factor Protocols

Transcription Factor Protocols PDF Author: Martin J. Tymms
Publisher: Springer Science & Business Media
ISBN: 159259686X
Category : Medical
Languages : en
Pages : 304

Get Book Here

Book Description
The effort to sequence the human genome is now moving toward a c- clusion. As all of the protein coding sequences are described, an increasing emphasis will be placed on understanding gene function and regulation. One important aspect of this analysis is the study of how transcription factors re- late transcriptional initiation by RNA polymerase II, which is responsible for transcribing nuclear genes encoding messenger RNAs. The initiation of Class II transcription is dependent upon transcription factors binding to DNA e- ments that include the core or basal promoter elements, proximal promoter elements, and distal enhancer elements. General initiation factors are involved in positioning RNA polymerase II on the core promoter, but the complex - teraction of these proteins and transcriptional activators binding to DNA e- ments outside the core promoter regulate the rate of transcriptional initiation. This initiation process appears to be a crucial step in the modulation of mRNA levels in response to developmental and environmental signals. Transcription Factor Protocols provides step-by-step procedures for key techniques that have been developed to study DNA sequences and the protein factors that regulate the transcription of protein encoding genes. This volume is aimed at providing researchers in the field with the well-detailed protocols that have been the hallmark of previous volumes of the Methods in Molecular TM Biology series.

Irreversible Electroporation

Irreversible Electroporation PDF Author: Boris Rubinsky
Publisher: Springer Science & Business Media
ISBN: 364205420X
Category : Technology & Engineering
Languages : en
Pages : 320

Get Book Here

Book Description
Non-thermal irreversible electroporation is a new minimally invasive surgical p- cedure with unique molecular selectivity attributes – in fact it may be considered the first clinical molecular surgery procedure. Non-thermal irreversible electro- ration is a molecular selective mode of cell ablation that employs brief electrical fields to produce nanoscale defects in the cell membrane, which can lead to cell death, without an effect on any of the other tissue molecules. The electrical fields can be produced through contact by insertion of electrode needles around the undesirable tissue and non-invasively by electromagnetic induction. This new - dition to the medical armamentarium requires the active involvement and is of interest to clinical physicians, medical researchers, mechanical engineers, che- cal engineers, electrical engineers, instrumentation designers, medical companies and many other fields and disciplines that were never exposed in their training to irreversible electroporation or to a similar concept. This edited book is designed to be a comprehensive introduction to the field of irreversible electroporation to those that were not exposed or trained in the field before and can also serve as a reference manual. Irreversible electroporation is broad and interdisciplinary. Therefore, we have made an attempt to cover every one of the various aspects of the field from an introductory basic level to state of the art.

Plant Cell Culture Protocols

Plant Cell Culture Protocols PDF Author: Robert D. Hall
Publisher: Springer Science & Business Media
ISBN: 1592595839
Category : Science
Languages : en
Pages : 422

Get Book Here

Book Description
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. Plant Cell Culture Protocols's 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.

Pichia Protocols

Pichia Protocols PDF Author: James M Cregg
Publisher: Springer Science & Business Media
ISBN: 1588294293
Category : Science
Languages : en
Pages : 553

Get Book Here

Book Description
This book focuses on recent developments of Pichia pastoris as a recombinant protein production system. Highlighted topics include a discussion on the use of fermentors to grow Pichia pastoris, information on the O- and N-linked glycosylation, methods for labeling Pichia pastoris expressed proteins for structural studies, and the introduction of mutations in Pichia pastoris genes by the methods of restriction enzyme-mediated integration (REMI). Each chapter presents cutting-edge and cornerstone protocols for utilizing P. pastoris as a model recomibinant protein production system. This volume fully updates and expands upon the first edition.

Connexin Methods and Protocols

Connexin Methods and Protocols PDF Author: Roberto Bruzzone
Publisher: Springer Science & Business Media
ISBN: 1592590438
Category : Science
Languages : en
Pages : 495

Get Book Here

Book Description
Direct cell–cell communication is a common property of multicellular organisms that is achieved through membrane channels which are organized in gap junctions. The protein subunits of these intercellular channels, the connexins, form a multigene family that has been investigated in great detail in recent years. It has now become clear that, in different tissues, connexins speak several languages that control specific cellular functions. This progress has been made possible by the availability of new molecular tools and the improvement of basic techniques for the study of membrane channels, as well as by the use of genetic approaches to study protein function in vivo. More important, connexins have gained visibility because mutations in some connexin genes have been found to be linked to human genetic disorders. Connexin Methods and Protocols presents in detail a collection of te- niques currently used to study the cellular and molecular biology of connexins and their physiological properties. The field of gap junctions and connexin research has always been characterized by a multidisciplinary approach c- bining morphology, biochemistry, biophysics, and cellular and molecular biology. This book provides a series of cutting-edge protocols and includes a large spectrum of practical methods that are available to investigate the fu- tion of connexin channels. Connexin Methods and Protocols is divided into three main parts.

Agrobacterium Protocols

Agrobacterium Protocols PDF Author: Kan Wang
Publisher: Springer Science & Business Media
ISBN: 1597451304
Category : Science
Languages : en
Pages : 474

Get Book Here

Book Description
Agrobacterium tumefaciens is a soil bacterium that for more than a century has been known as a pathogen causing the plant crown gall disease. Unlike many other pathogens, Agrobacterium has the ability to deliver DNA to plant cells and permanently alter the plant genome. The discovery of this unique feature 30 years ago has provided plant scientists with a powerful tool to genetically transform plants for both basic research purposes and for agricultural development. Compared to physical transformation methods such as particle bomba- ment or electroporation, Agrobacterium-mediated DNA delivery has a number of advantages. One of the features is its propensity to generate a single or a low copy number of integrated transgenes with defined ends. Integration of a single transgene copy into the plant genome is less likely to trigger “gene silencing” often associated with multiple gene insertions. When the first edition of Agrobacterium Protocols was published in 1995, only a handful of plants could be routinely transformed using Agrobacterium. Agrobacterium-mediated transformation is now commonly used to introduce DNA into many plant species, including monocotyledon crop species that were previously considered non-hosts for Agrobacterium. Most remarkable are recent developments indicating that Agrobacterium can also be used to deliver DNA to non-plant species including bacteria, fungi, and even mammalian cells.

Pseudomonas Methods and Protocols

Pseudomonas Methods and Protocols PDF Author: Alain Filloux
Publisher: Humana
ISBN: 9781493944163
Category : Science
Languages : en
Pages : 0

Get Book Here

Book Description
In Pseudomonas aeruginosa, expert researchers in the field detail many of the methods which are now commonly used to study this fascinating microorganism. Chapters include microbiological methods to high-throughput molecular techniques that have been developed over the last decade. 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, Pseudomonas aeruginosa aids in the continuing study of new and cutting edge findings.