RNA Isolation and Characterization Protocols

RNA Isolation and Characterization Protocols

Author: Ralph Rapley

Publisher: Humana Press

Published: 2013-08-18

Total Pages: 264

ISBN-13: 9781489942500

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Ribonucleic acids are central to cellular and molecular processes and perform vital functions in both structural and functional roles. RNA molecules form the bridge between the stable genetic information contained within DNA and enzymes and proteins that carry out much of the metabolism within the cell. Many of the sites of protein synthesis, the ribosomes within the cell, are composed of these ribonucleic acids as are the tRNA molecules that deliver the amino acid building blocks to the ribosomes. Of all the RNA species, the nucleic acid intermediate, messenger RNA, is a desirable source of material to biologists, since this reflects much of, what ultimately, is translated into enzymes and proteins. In order to determine the qualitative and quantitative changes in mRNA expression, a vast number of molecular biological techniques have been developed. Key molecular methods that provide the means to initially isolate and analyze RNA molecules are the focus of this volume. In putting together this collection of protocols, we have tried to provide techniques that are most applicable and widely used. In particular, there are a number of iso- tion techniques included that have been developed, modified, or adapted to enable extraction from a variety of cell types, organisms, or subcellular organelles. Successful isolation of intact RNA is an essential starting point for any sub- quent analysis. This is why we have aimed to make this section comprehensive. The analysis of RNA is the focus of the following chapters.


RNA Purification and Analysis

RNA Purification and Analysis

Author: Douglas T. Gjerde

Publisher: John Wiley & Sons

Published: 2009-07-10

Total Pages: 206

ISBN-13: 3527627200

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This first book on the market covers the many new and important RNA species discovered over the past five years, explaining current methods for the enrichment, separation and purification of these novel RNAs. Building up from general principles of RNA biochemistry and biophysics, this book addresses the practical aspects relevant to the laboratory researcher throughout, while discussing the performance and potential problems of the methods discussed. An appendix contains a glossary with the important terms and techniques used in RNA analysis. By explaining the basic and working principles of the methods, the book allows biochemists and molecular biologists to gain much more expertise than by simply repeating a pre-formulated protocol, enabling them to select the procedure and materials best suited to the RNA analysis task at hand. As a result, they will be able to develop new protocols where needed and optimize and fine-tune the general purpose standard protocols that come with the purification equipment and instrumentation.


RNA Methodologies

RNA Methodologies

Author: Robert E. Farrell Jr.

Publisher: Elsevier

Published: 2010-07-22

Total Pages: 794

ISBN-13: 0080454763

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This laboratory guide represents a growing collection of tried, tested and optimized laboratory protocols for the isolation and characterization of eukaryotic RNA, with lesser emphasis on the characterization of prokaryotic transcripts. Collectively the chapters work together to embellish the RNA story, each presenting clear take-home lessons, liberally incorporating flow charts, tables and graphs to facilitate learning and assist in the planning and implementation phases of a project.RNA Methodologies, 3rd edition includes approximately 30% new material, including chapters on the more recent technologies of RNA interference including: RNAi; Microarrays; Bioinformatics. It also includes new sections on: new and improved RT-PCR techniques; innovative 5' and 3' RACE techniques; subtractive PCR methods; methods for improving cDNA synthesis.* Author is a well-recognized expert in the field of RNA experimentation and founded Exon-Intron, a well-known biotechnology educational workshop center * Includes classic and contemporary techniques * Incorporates flow charts, tables, and graphs to facilitate learning and assist in the planning phases of projects


RNA Abundance Analysis

RNA Abundance Analysis

Author: Hailing Jin

Publisher: Humana Press

Published: 2016-05-01

Total Pages: 248

ISBN-13: 9781493959167

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This book examines a wide range of techniques on RNA extraction, detection, quantification, visualization, and genome-wide profiling, from conventional methods to state-of-the-art high throughput approaches.


Arabidopsis Protocols

Arabidopsis Protocols

Author: Jose J. Sanchez-Serrano

Publisher: Methods in Molecular Biology

Published: 2013-09-21

Total Pages: 754

ISBN-13:

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Arabidopsis Protocols, Third Edition compiles some of the most recent methodologies developed to exploit the Arabidopsis genome. These methodologies cover from the guided access to public resources, to genetic, cell biology, biochemical and physiological techniques, including both those that are widely used as well as those novel techniques likely to open up new avenues of knowledge in the future. In addition, considering the recent unparalleled progress of the “omics” tools in Arabidopsis, leading experts have contributed sections on genome, transcriptome, proteome, metabolome and other whole-system approaches. Arabidopsis thaliana is acknowledged as the most important plant model system by the scientific community and Arabidopsis research has fundamentally influenced our understanding of the basic biology and ecology of plants. 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, Arabidopsis Protocols, Third Edition seeks to serve both experienced researchers and beginners with its detailed methodologies on this burgeoning scientific field.


Techniques in Molecular Systematics and Evolution

Techniques in Molecular Systematics and Evolution

Author: Rob DeSalle

Publisher: Birkhäuser

Published: 2013-12-01

Total Pages: 414

ISBN-13: 3034881258

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The amount of information that can be obtained by using molecular techniques in evolution, systematics and ecology has increased exponentially over the last ten years. The need for more rapid and efficient methods of data acquisition and analysis is growing accordingly. This manual presents some of the most important techniques for data acquisition developed over the last years. The choice and justification of data analysis techniques is also an important and critical aspect of modern phylogenetic and evolutionary analysis and so a considerable part of this volume addresses this important subject. The book is mainly written for students and researchers from evolutionary biology in search for methods to acquire data, but also from molecular biology who might be looking for information on how data are analyzed in an evolutionary context. To aid the user, information on web-located sites is included wherever possible. Approaches that will push the amount of information which systematics will gather in the


PCR Protocols

PCR Protocols

Author: John M. S. Bartlett

Publisher: Springer Science & Business Media

Published: 2008-02-03

Total Pages: 1083

ISBN-13: 1592593844

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In this new edition, the editors have thoroughly updated and dramatically expanded the number of protocols to take advantage of the newest technologies used in all branches of research and clinical medicine today. These proven methods include real time PCR, SNP analysis, nested PCR, direct PCR, and long range PCR. Among the highlights are chapters on genome profiling by SAGE, differential display and chip technologies, the amplification of whole genome DNA by random degenerate oligonucleotide PCR, and the refinement of PCR methods for the analysis of fragmented DNA from fixed tissues. Each fully tested protocol is described in step-by-step detail by an established expert in the field and includes a background introduction outlining the principle behind the technique, equipment and reagent lists, tips on trouble shooting and avoiding known pitfalls, and, where needed, a discussion of the interpretation and use of results.


Guidelines for Molecular Analysis in Archive Tissues

Guidelines for Molecular Analysis in Archive Tissues

Author: Giorgio Stanta

Publisher: Springer Science & Business Media

Published: 2011-06-06

Total Pages: 301

ISBN-13: 3642178901

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A huge amount of fixed and paraffin-embedded tissue is stored in every hospital. This is very precious material that can be used for translational research and for diagnostics. The molecular methods employed for analysis of these tissues are similar to the usual molecular biology and proteomics methods, but reliable results can be obtained only if specific steps are followed with great care. This book provides detailed and precise guidelines for molecular analysis of archive tissues and will serve as an invaluable aid for researchers and pathologists involved in translational research and diagnostics. Clear notes and explanations are included to simplify use of the protocols for the less experienced. The authors are a group of acknowledged experts who have developed the described methods and validated them within the European project "Archive Tissues: Improving Molecular Medicine Research and Clinical Practice - IMPACTS", which has involved 21 leading institutions in 11 countries.


A Laboratory Guide to RNA

A Laboratory Guide to RNA

Author: Paul A. Krieg

Publisher: John Wiley & Sons

Published: 1996-08-15

Total Pages: 470

ISBN-13: 9780471125365

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Here is the most complete guide available to the isolation, analysis, and synthesis of RNA. It covers everything researchers and laboratory workers need to know about the study of gene expression via RNA analysis-from the theory behind the methods, to actual problem-solving techniques. Step-by-step protocols are presented for each method. A careful presentation of the experimental formalities of these protocols enables specialists and nonspecialists alike to implement the methods easily in the laboratory. Each protocol is accompanied by the theoretical background underlying the experimental procedure and most chapters contain illustrations of typical results and troubleshooting tips. A Laboratory Guide to RNA offers a straightforward detailed account of experimental procedures, ranging from the isolation of RNA from a variety of cell and tissue types, detection analysis, and quantitation using a range of strategies, to large- and small-scale synthesis of RNA. This unique guide not only covers established procedures such as RNA blotting and nuclease protection, but also the latest protocols for quantitative PCR and differential display. Protocols addressing in situ hybridization are highlighted in an eight-page, full-color section that illustrates the power of the technique for detection of gene expression in tissues and whole organisms. Featuring contributions from leading research laboratories and the biotechnology field, A Laboratory Guide to RNA: Isolation, Analysis, and Synthesis provides all the methods required for RNA analysis. It is the ideal laboratory guide for research scientists, graduate students, and lab personnel who need a solid reference on the analysis of gene expression at the RNA level.