Nuclease Methods and Protocols

Nuclease Methods and Protocols PDF Author: Catherine H. Schein
Publisher: Springer Science & Business Media
ISBN: 1592592333
Category : Science
Languages : en
Pages : 521

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Book Description
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.

Developmental Biology Protocols

Developmental Biology Protocols PDF Author: Rocky S. Tuan
Publisher: Springer Science & Business Media
ISBN: 1592590659
Category : Science
Languages : en
Pages : 537

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Book Description
Developmental biology is one of the most exciting and fast-growing fields today. In part, this is so because the subject matter deals with the innately fascinating biological events—changes in form, structure, and function of the org- ism. The other reason for much of the excitement in developmental biology is that the field has truly become the unifying melting pot of biology, and provides a framework that integrates anatomy, physiology, genetics, biochemistry, and cellular and mole- lar biology, as well as evolutionary biology. No longer is the study of embryonic development merely “embryology.” In fact, development biology has produced - portant paradigms for both basic and clinical biomedical sciences alike. Although modern developmental biology has its roots in “experimental emb- ology” and the even more classical “chemical embryology,” the recent explosive and remarkable advances in developmental biology are critically linked to the advent of the “cellular and molecular biology revolution.” The impressive arsenal of expe- mental and analytical tools derived from cell and molecular biology, which promise to continue to expand, together with the exponentially developing sophistication in fu- tional imaging and information technologies, guarantee that the study of the devel- ing embryo will contribute one of the most captivating areas of biological research in the next millennium.

Nuclease Methods and Protocols

Nuclease Methods and Protocols PDF Author: Catherine H. Schein
Publisher: Springer Science & Business Media
ISBN: 1592592333
Category : Science
Languages : en
Pages : 521

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Book Description
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.

Transgenic Mouse Methods and Protocols

Transgenic Mouse Methods and Protocols PDF Author: Marten H. Hofker
Publisher: Springer Science & Business Media
ISBN: 1592593402
Category : Science
Languages : en
Pages : 382

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Book Description
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.

Advanced Protocols for Animal Transgenesis

Advanced Protocols for Animal Transgenesis PDF Author: Shirley Pease
Publisher: Springer Nature
ISBN: 3642207928
Category : Science
Languages : en
Pages : 671

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Book Description
This laboratory manual, published in cooperation with the International Society for Transgenic Technology (ISTT), provides almost all current methods that can be applied to the creation and analysis of genetically modified animals. The chapters have been contributed by leading scientists who are actively using the technology in their laboratories. Based on their first-hand experience the authors also provide helpful notes and troubleshooting sections. Topics range from standard techniques, such as pronuclear microinjection of DNA, to more sophisticated and modern methods, such as the derivation and establishment of embryonic stem (ES) cell lines, with defined inhibitors in cell culture medium. In addition, related topics with relevance to the field are addressed, including global web-based resources, legal issues, colony management, shipment of mice and embryos, and the three R’s: refinement, reduction and replacement.

Gene Isolation and Mapping Protocols

Gene Isolation and Mapping Protocols PDF Author: Jacqueline Boultwood
Publisher: Springer Science & Business Media
ISBN: 1592595545
Category : Science
Languages : en
Pages : 323

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Book Description
An unprecedented collection of all the most up-to-date techniques for gene isolation and mapping, including the latest methods for gene characterization using database analyses. This collection of thoroughly tested recipes also includes chapters for the computational analysis of novel cDNA sequences with up-to-the-minute information on basic sequence analysis, sequence similarity searches, exon detection and similarity searches, and the prediction of gene function. Its state-of-the-art methods constitute indispensable tools for all scientists engaged in the search for specific disease genes, or in the general advancement of the human genome project.

Microarrays

Microarrays PDF Author: Jang B. Rampal
Publisher: Springer Science & Business Media
ISBN: 1588295893
Category : Medical
Languages : en
Pages : 923

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Book Description
Microarray Technology, Volumes 1 and 2, present information in designing and fabricating arrays and binding studies with biological analytes while providing the reader with a broad description of microarray technology tools and their potential applications. The first volume deals with methods and protocols for the preparation of microarrays. The second volume details applications and data analysis, which is important in analyzing the enormous data coming out of microarray experiments. Among the topics discussed in Volume 1: Synthesis Methods, are matrices in the synthesis of microarrays, array optimization processes, array-based comparative genomic hybridization, 60-mer oligonucleotide probes, bifunctional reagents NTMTA and NTPAC, and high density arrays using digital microarray synthesis platforms. Other topics include multiplex ligation-dependent probe amplification (MLPA), hybridization conditions in situ-synthesized oligo arrays, peptide arrays, high density replication tools (HDRT), protocols for the quantification of oligo hybridization, glyco-bead arrays, and an investigation into the emerging nano technology. Microarray Technology, Volumes 1 and 2, provide ample information to all levels of scientists from novice to those intimately familiar with array technology.

High Content Screening

High Content Screening PDF Author: D. Lansing Taylor
Publisher: Springer Science & Business Media
ISBN: 1597452173
Category : Science
Languages : en
Pages : 437

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Book Description
There has always been some tension between proponents of hypothesis-driven and discovery-driven research in the broad field of life sciences. Academic research has been primarily focused on hypothesis-driven research. However, the success of the human genome project, a discovery-driven research approach, has opened the door to adding other types of discovery-driven research to a continuum of research approaches. In contrast, drug discovery research in the pharmaceutical industry has embraced discovery-driven research for many years. A good example has been the discovery of active compounds from large chemical libraries, through screening campaigns. The success of the human genome project has also demonstrated the need for both academic researchers and industrial researchers to now understand the functions of genes and gene products. The cell is the basic unit of life and it has been at the cellular level where function can be demonstrated most cost-effectively and rapidly. High content screening (HCS) was developed by Cellomics Inc. in the mid-1990s to address the need for a platform that could be used in the discovery-driven research and development required to understand the functions of genes and gene products at the level of the cell.

Encyclopedia of Genetics

Encyclopedia of Genetics PDF Author: Eric C.R. Reeve
Publisher: Routledge
ISBN: 1134263503
Category : Reference
Languages : en
Pages : 985

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Book Description
The Encyclopedia includes 125 entries, beginning with the origins of genetics including historical background on the work of Gregor Mendel and Charles Darwin, and progressing to the structure of DNA and modern theories such as selfish genes. All branches of genetics are covered, including the genetics of bacteria, viruses, insects, animals and plants, as well as humans. Important topical issues such as the human genome project, bioethics, the law and genetics, genetic disorders, GM crops, and the use of transgenic animals for food and pharmaceutical products are fully surveyed. A section on techniques and biotechnology includes modern methods of analysis, from DNA fingerprinting to the new science of bioinformatics. The articles, all written by specialists, are largely non-mathematical and progress from general concepts to deeper understanding. Each essay is fully referenced, with suggestions for further reading. The text is supplemented by extensive illustrations, tables and a color plate section. The Encyclopedia of Genetics will be a valuable companion for all those working or studying in the various fields of genetical research, and a fascinating reference for all readers with a basic background in biology. Also includes color inserts.

Protocols in Human Molecular Genetics

Protocols in Human Molecular Genetics PDF Author: Christopher G. Mathew
Publisher: Springer Science & Business Media
ISBN: 1592594964
Category : Medical
Languages : en
Pages : 461

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Book Description
Extraordinary advances have been made in the field of human molecular genetics during the past five years. The ability to amplify a specific region ofDNA a millionfold in a few hours using the polymerase chain reaction has led to the rapid identification of mutations in human disease and of DNA sequence polymorphisms on every human chro- some. DNA fragments of up to 1 megabase in length can now be resolved by pulsed-field gel electrophoresis to create long-range physical maps of important regions of the genome, and can be cloned in the form of yeast artificial chromosomes. The discovery of highly variable "minisatellite" DNA sequences has led to the development of DNA fingerprinting. The application of these techniques to the study of the human genome has culminated in major advances such as the cloning of the cystic fibrosis gene, the construction of genetic linkage maps of each human chro- some, the mapping of many genes responsible for human inherited d- orders, genetic fingerprinting of forensic specimens, and the detection of mutations involved in the development of human tumors. Although many of the new techniques in molecular genetics can be learned relatively easily, it is often difficult for a researcher to obtain all of the relevant information necessary for getting up a technique and applying it successfully. The information available in the research lite- ture often lacks the depth of description that the new user requires.

Chromosome Analysis Protocols

Chromosome Analysis Protocols PDF Author: John R. Gosden
Publisher: Springer Science & Business Media
ISBN: 1592595162
Category : Medical
Languages : en
Pages : 508

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Book Description
Chromosomes, as the genetic vehicles, provide the basic material for a large proportion of genetic investigations, from the construction of gene maps and models of chromosome organization, to the inves tigation of gene function and dysfunction. The study of chromosomes has developed in parallel with other aspects of molecular genetics, beginning with the first preparations of chromosomes from animal cells, through the development of banding techniques, which permitted the unequivocal identification of each chromosome in a karyotype, to the present analytical methods of molecular cytogenetics. Although some of these techniques have been in use for many years, and can be learned relatively easily, most published scientific reports—as a result of pressure on space from editors, and the response to that pressure by authors—contain little in the way of technical detail, and thus are rarely adequate for a researcher hoping to find all the necessary information to embark on a method from scratch. A new user needs not only a detailed description of the methods, but also some help with problem solving and sorting out the difficulties en countered in handling any biological system. This was the require ment to which the series Methods in Molecular Biology is addressed, and Chromosome Analysis Protocols forms a part of this series.