Nuclease Methods And Protocols

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Nuclease Methods and Protocols

Catherine H. Schein
Nuclease Methods and Protocols Book PDF

Author : Catherine H. Schein
Publisher : Springer Science & Business Media
Page : 521 pages
File Size : 41,6 Mb
Release : 2008-02-03
Category : Science
ISBN : 9781592592333

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Nuclease Methods and Protocols by Catherine H. Schein Pdf

Nucleases, enzymes that restructure or degrade nucleic acid polymers, are vital to the control of every area of metabolism. They range from “housekeeping” enzymes with broad substrate ranges to extremely specific tools (1). Many types of nucleases are used in lab protocols, and their commercial and clinical uses are expanding. The purpose of Nuclease Methods and Protocols is to introduce the reader to some we- characterized protein nucleases, and the methods used to determine their activity, structure, interaction with other molecules, and physiological role. Each chapter begins with a mini-review on a specific nuclease or a nuclease-related theme. Although many chapters cover several topics, they were arbitrarily divided into five parts: Part I, “Characterizing Nuclease Activity,” includes protocols and assays to determine general (processive, distributive) or specific mechanisms. Methods to assay nuclease products, identify cloned nucleases, and determine their physiological role are also included here. Part II, “Inhibitors and Activators of Nucleases,” summarizes assays for measuring the effects of other proteins and small molecules. Many of these inhibitors have clinical relevance. Part III, “Relating Nuclease Structure and Function,” provides an overview of methods to determine or model the 3-D structure of nucleases and their complexes with substrates and inhibitors. A 3-D structure can greatly aid the rational design of nucleases and inhibitors for specific purposes. Part IV, “Nucleases in the Clinic,” summarizes assays and protocols suitable for use with t- sues and for nuclease based therapeutics.

Nuclease Methods and Protocols

Catherine H. Schein
Nuclease Methods and Protocols Book PDF

Author : Catherine H. Schein
Publisher : Humana Press
Page : 525 pages
File Size : 52,9 Mb
Release : 2001-01-30
Category : Science
ISBN : 0896036790

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Nuclease Methods and Protocols by Catherine H. Schein Pdf

Nucleases, enzymes that restructure or degrade nucleic acid polymers, are vital to the control of every area of metabolism. They range from “housekeeping” enzymes with broad substrate ranges to extremely specific tools (1). Many types of nucleases are used in lab protocols, and their commercial and clinical uses are expanding. The purpose of Nuclease Methods and Protocols is to introduce the reader to some we- characterized protein nucleases, and the methods used to determine their activity, structure, interaction with other molecules, and physiological role. Each chapter begins with a mini-review on a specific nuclease or a nuclease-related theme. Although many chapters cover several topics, they were arbitrarily divided into five parts: Part I, “Characterizing Nuclease Activity,” includes protocols and assays to determine general (processive, distributive) or specific mechanisms. Methods to assay nuclease products, identify cloned nucleases, and determine their physiological role are also included here. Part II, “Inhibitors and Activators of Nucleases,” summarizes assays for measuring the effects of other proteins and small molecules. Many of these inhibitors have clinical relevance. Part III, “Relating Nuclease Structure and Function,” provides an overview of methods to determine or model the 3-D structure of nucleases and their complexes with substrates and inhibitors. A 3-D structure can greatly aid the rational design of nucleases and inhibitors for specific purposes. Part IV, “Nucleases in the Clinic,” summarizes assays and protocols suitable for use with t- sues and for nuclease based therapeutics.

Biotechnologies for Plant Mutation Breeding

Joanna Jankowicz-Cieslak,Thomas H. Tai,Jochen Kumlehn,Bradley J. Till
Biotechnologies for Plant Mutation Breeding Book PDF

Author : Joanna Jankowicz-Cieslak,Thomas H. Tai,Jochen Kumlehn,Bradley J. Till
Publisher : Springer
Page : 343 pages
File Size : 43,8 Mb
Release : 2016-12-08
Category : Science
ISBN : 9783319450216

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Biotechnologies for Plant Mutation Breeding by Joanna Jankowicz-Cieslak,Thomas H. Tai,Jochen Kumlehn,Bradley J. Till Pdf

This book is open access under a CC BY-NC 2.5 license. This book offers 19 detailed protocols on the use of induced mutations in crop breeding and functional genomics studies, which cover topics including chemical and physical mutagenesis, phenotypic screening methods, traditional TILLING and TILLING by sequencing, doubled haploidy, targeted genome editing, and low-cost methods for the molecular characterization of mutant plants that are suitable for laboratories in developing countries. The collection of protocols equips users with the techniques they need in order to start a program on mutation breeding or functional genomics using both forward and reverse-genetic approaches. Methods are provided for seed and vegetatively propagated crops (e.g. banana, barley, cassava, jatropha, rice) and can be adapted for use in other species.

Connexin Methods and Protocols

Roberto Bruzzone,Christian Giaume
Connexin Methods and Protocols Book PDF

Author : Roberto Bruzzone,Christian Giaume
Publisher : Springer Science & Business Media
Page : 495 pages
File Size : 40,8 Mb
Release : 2008-02-05
Category : Science
ISBN : 9781592590438

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Connexin Methods and Protocols by Roberto Bruzzone,Christian Giaume Pdf

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.

RNA

Henrik Nielsen
RNA Book PDF

Author : Henrik Nielsen
Publisher : Humana Press
Page : 329 pages
File Size : 53,5 Mb
Release : 2010-12-10
Category : Medical
ISBN : 1588299139

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RNA by Henrik Nielsen Pdf

Recent insight into the transcripts generated from the mammalian genome (i.e. the transcriptome) has revealed that transcription is a far more complex phenomenon than previously thought. In RNA: Methods and Protocols, expert researchers provide the procedures and methods used to describe the structure of messenger RNAs and non-coding RNAs that are transcribed by RNA polymerase II as the immediate gene products in mammalian cells. Focused on the structure of the RNA products of “gene X” and the mapping of proteins associated with these RNAs, the volume presents appropriate information for non-specialists in RNA biology. Written in the highly successful Methods in Molecular BiologyTM series format, many chapters contain 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. Comprehensive and practical, RNA: Methods and Protocols views the transcriptional landscape with an appreciation for the role that proteins play in the processing and interpretation of genetic information in an attempt to further our crucial knowledge of the many products and sophisticated regulatory networks that result from it.

CRISPR

Magnus Lundgren,Emmanuelle Charpentier,Peter C. Fineran
CRISPR Book PDF

Author : Magnus Lundgren,Emmanuelle Charpentier,Peter C. Fineran
Publisher : Humana
Page : 0 pages
File Size : 46,8 Mb
Release : 2016-10-09
Category : Medical
ISBN : 1493954288

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CRISPR by Magnus Lundgren,Emmanuelle Charpentier,Peter C. Fineran Pdf

This volume presents a list of cutting-edge protocols for the study of CRISPR-Cas defense systems and their applications at the genomic, genetic, biochemical and structural levels. CRISPR: Methods and Protocols guides readers through techniques that have been developed specifically for the analysis of CRISPR-Cas and techniques adapted from standard protocols of DNA, RNA and protein biology. 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, CRISPR: Methods and Protocols provides a broad list of tools and techniques to study the interdisciplinary aspects of the prokaryotic CRISPR-Cas defense systems.

Liposome Methods and Protocols

Subhash C. Basu,Manju Basu
Liposome Methods and Protocols Book PDF

Author : Subhash C. Basu,Manju Basu
Publisher : Springer Science & Business Media
Page : 250 pages
File Size : 51,5 Mb
Release : 2008-02-04
Category : Medical
ISBN : 9781592591756

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Liposome Methods and Protocols by Subhash C. Basu,Manju Basu Pdf

In vitro utilization of liposomes is now recognized as a powerful tool in many bioscience investigations and their associated clinical studies, e.g., liposomes in drug targeting; liposomes in gene transport across plasma and nuclear membranes; liposomes in enzyme therapy in patients with genetic disorders. However, before these areas can be effectively explored, many basic areas in liposome research require elucidation, including: (a) attachment of liposomes to cell surfaces; (b) permeation of liposomes through the plasma membranes; and (c) stability of liposomes in cell or nuclear matrices. None of these areas have been exhaustively explored and liposome researchers have ample opportunities to contribute to our knowledge. The aim of Liposome Methods and Protocols is to bring together a wide range of detailed laboratory protocols covering different aspects of liposome biology in order to assist researchers in those rapidly advancing medical fields mentioned earlier. With this goal in mind, in each protocol chapter we have detailed the materials to be used, followed by a step-by-step protocol. The Notes section of each protocol is also certain to prove particularly useful, since the authors include troubleshooting tips straight from their benchtops, valuable information that is seldom given in restricted methods sections of standard research journals. For this reason we feel that the book will prove especially useful for all researchers in the liposome field.

Transgenic Mouse Methods and Protocols

Marten H. Hofker,Jan van Deursen
Transgenic Mouse Methods and Protocols Book PDF

Author : Marten H. Hofker,Jan van Deursen
Publisher : Springer Science & Business Media
Page : 382 pages
File Size : 47,8 Mb
Release : 2008-02-04
Category : Science
ISBN : 9781592593408

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Transgenic Mouse Methods and Protocols by Marten H. Hofker,Jan van Deursen Pdf

Marten Hofker and Jan van Deursen have assembled a multidisciplinary collection of readily reproducible methods for working with mice, and particularlyfor generating mouse models that will enable us to better understand gene function. Described in step-by-step detail by highly experienced investigators, these proven techniques include new methods for conditional, induced knockout, and transgenic mice, as well as for working with mice in such important research areas as immunology, cancer, and atherosclerosis. Such alternative strategies as random mutagenesis and viral gene transduction for studying gene function in the mouse are also presented.

Steroid Receptor Methods

Benjamin A. Lieberman
Steroid Receptor Methods Book PDF

Author : Benjamin A. Lieberman
Publisher : Springer Science & Business Media
Page : 377 pages
File Size : 48,6 Mb
Release : 2001-08-10
Category : Medical
ISBN : 9780896037540

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Steroid Receptor Methods by Benjamin A. Lieberman Pdf

A distinguished team of principal investigators and their associates describe in step-by-step detail a cross-section of the latest research techniques available for studying the endocrine system. As a basis for sophisticated biochemical analysis of receptor properties, the contributors provide methods for the production and purification of a variety of receptors, including progesterone, glucocorticoid, and androgen. Other protocols allow the reader to experiment with DNA binding characteristics, hormone binding assays, and the use of combinatorial chemistry for drug discovery. A series of novel methods utilizing the latest advances in immunochemistry, yeast two-hybrid screening, and fluorescence are included for the detection and analysis of a variety of cellular proteins that influence steroid receptor effectiveness.

Proteoglycan Protocols

Renato V. Iozzo
Proteoglycan Protocols Book PDF

Author : Renato V. Iozzo
Publisher : Springer Science & Business Media
Page : 547 pages
File Size : 50,5 Mb
Release : 2008-02-02
Category : Science
ISBN : 9781592592098

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Proteoglycan Protocols by Renato V. Iozzo Pdf

Proteoglycans are some of the most elaborate macromolecules of mammalian and lower organisms. The covalent attachment of at least five types of glycosami- glycan side chains to more than forty individual protein cores makes these molecules quite complex and endows them with a multitude of biological functions. Proteoglycan Protocols offers a comprehensive and up-to-date collection of prepa- tive and analytical methods for the in-depth analysis of proteoglycans. Featuring st- by-step detailed protocols, this book will enable both novice and experienced researchers to isolate intact proteoglycans from tissues and cultured cells, to establish the composition of their carbohydrate moieties, to generate strategies for prokaryotic and eukaryotic expression, to utilize methods for the suppression of specific proteoglycan gene expression and for the detection of mutant cells and degradation products, and to study specific interactions between proteoglycans and extracellular matrix proteins as well as growth factors and their receptors. The readers will find concise, yet comprehensive techniques carefully drafted by leading experts in the field. Each chapter commences with a general Introduction, followed by a detailed Materials section, and an easy-to-follow Methods section. An asset of each chapter is the extensive notation that includes troubleshooting tips and practical considerations that are often lacking in formal methodology papers. The reader will find this section most valuable because it is clearly provided by experienced scientists who have first-hand knowledge of the techniques they outline. In addition, most of the chapters are well illustrated with examples of typical data generated with each method.

Genomics Protocols

Michael P. Starkey,Ramnath Elaswarapu
Genomics Protocols Book PDF

Author : Michael P. Starkey,Ramnath Elaswarapu
Publisher : Springer Science & Business Media
Page : 537 pages
File Size : 55,6 Mb
Release : 2008-02-03
Category : Medical
ISBN : 9781592592357

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Genomics Protocols by Michael P. Starkey,Ramnath Elaswarapu Pdf

We must unashamedly admit that a large part of the motivation for editing Genomics Protocols was selfish. The possibility of assembling in a single volume a unique and comprehensive collection of complete protocols, relevant to our work and the work of our colleagues, was too good an opportunity to miss. We are pleased to report, however, that the outcome is something of use not only to those who are experienced practitioners in the genomics field, but is also valuable to the larger community of researchers who have recognized the potential of genomics research and may themselves be beginning to explore the technologies involved. Some of the techniques described in Genomics Protocols are clearly not restricted to the genomics field; indeed, a prerequisite for many procedures in this discipline is that they require an extremely high throughput, beyond the scope of the average investigator. However, what we have endeavored here to achieve is both to compile a collection of procedures concerned with geno- scale investigations and to incorporate the key components of “bottom-up” and “top-down” approaches to gene finding. The technologies described extend from those traditionally recognized as coming under the genomics umbrella, touch on proteomics (the study of the expressed protein complement of the genome), through to early therapeutic approaches utilizing the potential of genome programs via gene therapy (Chapters 27–30).

Gene Knockout Protocols

Martin J. Tymms,Ismail Kola
Gene Knockout Protocols Book PDF

Author : Martin J. Tymms,Ismail Kola
Publisher : Springer Science & Business Media
Page : 434 pages
File Size : 51,8 Mb
Release : 2008-02-03
Category : Medical
ISBN : 9781592592203

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Gene Knockout Protocols by Martin J. Tymms,Ismail Kola Pdf

As the major task of sequencing the human genome is near completion and full complement of human genes are catalogued, attention will be focused on the ultimate goal: to understand the normal biological functions of these genes, and how alterations lead to disease states. In this task there is a severe limitation in working with human material, but the mouse has been adopted as the favored animal model because of the available genetic resources and the highly conserved gene conservation linkage organization. In just of ten years since the first gene-targeting experiments were p- formed in embryonic stem (ES) cells and mutations transmitted through the mouse germline, more than a thousand mouse strains have been created. These achievements have been made possible by pioneering work that showed that ES cells derived from preimplantation mouse embryos could be cultured for prolonged periods without differentiation in culture, and that homologous rec- bination between targeting constructs and endogenous DNA occurred at a f- quency sufficient for recombinants to be isolated. In the next few years the mouse genome will be systematically altered, and the techniques for achi- ing manipulations are constantly being streamlined and improved.

PCR Protocols

Bruce A. White
PCR Protocols Book PDF

Author : Bruce A. White
Publisher : Springer Science & Business Media
Page : 397 pages
File Size : 41,5 Mb
Release : 2008-02-02
Category : Science
ISBN : 9781592595020

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PCR Protocols by Bruce A. White Pdf

PCR has been successfully utilized in every facet of basic, cli- cal, and applied studies of the life sciences, and the impact that PCR has had on life science research is already staggering. C- comitant with the essentially universal use of PCR has been the creative and explosive development of a wide range of PCR-based techniques and applications. These increasingly numerous pro- cols have each had the general effect of facilitating and acceler- ing research. Because PCR technology is relatively easy and inexpensive, PCR applications are well within the reach of every research lab. In this sense, PCR has become the "equalizer" between "small" and "big" labs, since its use makes certain projects, especially those related to molecular cloning, now far more feasible for the small lab with a modest budget. This new volume on PCR Protocols does not attempt the impossible task of representing all PCR-based protocols. Rather, it presents a range of protocols, both analytical and preparative, that provide a solid base of knowledge on the use of PCR in many c- mon research problems. The first six chapters provide some basic information on how to get started. Chapters 7-19 represent primarily analytical uses of PCR, both for simple DNA and RNA detection, as well as for more complex analyses of nucleic acid (e. g. , DNA footprin ting, RNA splice site localization). The remaining chapters represent "synthetic," or preparative, uses of PCR.

Adipose Tissue Protocols

Gérard Ailhaud
Adipose Tissue Protocols Book PDF

Author : Gérard Ailhaud
Publisher : Springer Science & Business Media
Page : 332 pages
File Size : 47,7 Mb
Release : 2008-02-03
Category : Science
ISBN : 9781592592319

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Adipose Tissue Protocols by Gérard Ailhaud Pdf

Adipose tissue is recognized to be exquisitely sensitive to hormone action, and is also now recognized as a secretory and endocrine organ required for reproduction and good health. Adipocytes are “smart” cells able within the tissue to communicate with surrounding cells, but also with various organs, particularly via leptin acting on the central nervous system. Brown adipose tissue (BAT) and white adipose tissue (WAT) are known to be distinct tissues, whereas the heterogeneity of WAT depots is well est- lished. Unfortunately, excess WAT leads to obesity, which is the most common health problem in industrialized countries. Therefore, from both a scientific and a technical point of view, the time has come to create a survey of adipose tissues and their neglected adipocytes. In Adipose Tissue Protocols, I have attempted to gather together chapters from all areas of adipose tissue research—from in vivo to in vitro studies—and to provide methods covering a wide variety of techniques, including the choice of adipose tissue depot and of morphological techniques for the study of BAT and WAT; the isolation, subcellular fractionation, and transfection of adipocytes where the low density of these cells must be taken into account; assays of nutrient and ion fluxes and the metabolic aspects of nutrient uptake; assays of lipid-related enzymes; biopsies and quantification of lipid-related mRNAs; cultures of adipose precursor cells from WAT and BAT of various species, including human tissue; measurements of adipose secretory products; and assessment of WAT metabolism in vivo.

Malaria Methods and Protocols

Denise L. Doolan
Malaria Methods and Protocols Book PDF

Author : Denise L. Doolan
Publisher : Springer Science & Business Media
Page : 606 pages
File Size : 52,9 Mb
Release : 2008-02-02
Category : Medical
ISBN : 9781592592715

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Malaria Methods and Protocols by Denise L. Doolan Pdf

The Plasmodium spp. parasite was identified as the causative agent of malaria in 1880, and the mosquito was identified as the vector in 1897. Despite subsequent efforts focused on the epidemiology, cell biology, immunology, molecular biology, and clinical manifestations of malaria and the Plasmodium parasite, there is still no licensed vaccine for the prevention of malaria. Physical barriers (bed nets, window screens) and chemical prevention methods (insecticides and mosquito repellents) intended to interfere with the transmission of the disease are not highly effective, and the profile of resistance of the parasite to chemoprophylactic and chemotherapeutic agents is increasing. The dawn of the new millennium has seen a resurgence of interest in the disease by government and philanthropic organizations, but we are still faced with compl- ities of the parasite, the host, and the vector, and the interactions among them. Malaria Methods and Protocols offers a comprehensive collection of protocols describing conventional and state-of-the-art techniques for the study of malaria, as well as associated theory and potential problems, written by experts in the field. The major themes reflected here include assessing the risk of infection and severity of disease, laboratory models, diagnosis and typing, molecular biology techniques, immunological techniques, cell biology techniques, and field applications.