Differential Display Methods and Protocols
Differential Display Methods and Protocols

Author: Peng Liang

Publisher: Springer Science & Business Media

Published: 2008-02-04

Total Pages: 321

ISBN-13: 1592599680

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Since the first edition of this book dedicated to differential display (DD) technology was published in 1997, we have witnessed an explosive interest in studying differential gene expression. The gene-hunting euphoria was initially powered by the invention of DD, which was gradually overtaken by DNA microarray technology in recent years. Then why is there still the need for second edition of this DD book? First of all, DD still enjoys a substantial lead over DNA microarrays in the ISI citation data (see Table 1), despite the h- dreds of millions of dollars spent each year on arrays. This may come as a surprise to many, but to us it implies that many of the DNA microarray studies went unpublished owing to their unfulfilled promises (1). Second, unlike DNA microarrays, DD is an “open”-ended gene discovery method that does not depend on prior genome sequence information of the organism being studied. As such, DD is applicable to the study of all living organisms—from bacteria, fungi, insects, fish, plants, to mammals—even when their genomes are not sequenced. Second, DD is more accessible technically and financially to most cost-conscious “cottage-industry” academic laboratories. So clearly DD still has its unique place in the modern molecular biological toolbox for gene expression analysis.

Differential-Display Reverse Transcription-PCR (DDRT-PCR)
Differential-Display Reverse Transcription-PCR (DDRT-PCR)

Author: Sergio Colonna-Romano

Publisher: Springer Science & Business Media

Published: 2012-12-06

Total Pages: 135

ISBN-13: 3642804543

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Identification of differentially expressed genes is one of the major challenges in molecular biology. Several techniques allow the cloning of such sequences. However, methods such as RNA subtraction or differential hybridization are time-consuming and require large amounts of mRNA. Recently, a new approach has successfully been developed: Differential-Display Reverse Transcription-PCR (DDRT-PCR). This technique has been proven to be highly effective in identifying sequences that are differentially expressed in various cell types. The most striking advantage is, however, that only nanograms of total RNA are sufficient. Thus every mRNA species expressed in the cell system can be investigated, even those present at very low levels.

Differential Display Methods and Protocols
Differential Display Methods and Protocols

Author: Peng Liang

Publisher: Humana Press

Published: 2005-10-21

Total Pages: 320

ISBN-13: 9781588293381

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Since the first edition of this book dedicated to differential display (DD) technology was published in 1997, we have witnessed an explosive interest in studying differential gene expression. The gene-hunting euphoria was initially powered by the invention of DD, which was gradually overtaken by DNA microarray technology in recent years. Then why is there still the need for second edition of this DD book? First of all, DD still enjoys a substantial lead over DNA microarrays in the ISI citation data (see Table 1), despite the h- dreds of millions of dollars spent each year on arrays. This may come as a surprise to many, but to us it implies that many of the DNA microarray studies went unpublished owing to their unfulfilled promises (1). Second, unlike DNA microarrays, DD is an “open”-ended gene discovery method that does not depend on prior genome sequence information of the organism being studied. As such, DD is applicable to the study of all living organisms—from bacteria, fungi, insects, fish, plants, to mammals—even when their genomes are not sequenced. Second, DD is more accessible technically and financially to most cost-conscious “cottage-industry” academic laboratories. So clearly DD still has its unique place in the modern molecular biological toolbox for gene expression analysis.

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.

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.

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

Cancer Genomics and Proteomics
Cancer Genomics and Proteomics

Author: Paul B. Fisher

Publisher: Humana Press

Published: 2007-09-27

Total Pages: 358

ISBN-13: 9781588295040

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Cancer Genomics and Proteomics provides a compendium of techniques and applications in gene identification and function. The approaches described in detail are state-of-the art and can be tailored to individual ongoing or planned research projects. This volume is a valuable laboratory resource for designing experiments to identify and analyze genes that are relevant to complex biological phenomena.

Molecular Embryology
Molecular Embryology

Author: Paul T. Sharpe

Publisher: Springer Science & Business Media

Published: 2008-02-02

Total Pages: 752

ISBN-13: 1592592708

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Most people have some interest in embryos; this probably results, in part, from their interest in understanding the biological origins of themselves and their offspring and, increasingly, concerns about how environmental change such as pollution might affect human development. Obviously, et- cal considerations preclude experimental studies of human embryos and, c- sequently, the developmental biologist has turned to other species to examine this process. Fortunately, the most significant conclusion to be drawn from the experimental embryology of the last two decades is the manner in which orthologous or closely related molecules are deployed to mediate similar - velopmental processes in both vertebrates and invertebrates. The molecular mechanisms regulating processes fundamental to most animals, such as axial patterning or axon guidance, are frequently conserved during evolution. (It is now widely believed that the differences between phyla and classes are the result of new genes, arising mostly by duplication and divergence of extant sequences, regulating the appearance of derived characters. ) Other vertebrates are obviously most likely to use the same devel- mental mechanisms as humans and, within the vertebrate subphylum, the - parent degree of conservation of developmental mechanism is considerable. It has long been recognized that particular vertebrate species offer either d- tinct advantages in investigating particular stages of development or are - pecially amenable to particular manipulations. No single animal can provide all the answers because not all types of experiments can be carried out on a single species.

Molecular Biomethods Handbook
Molecular Biomethods Handbook

Author: John M. Walker

Publisher: Springer Science & Business Media

Published: 2008-11-04

Total Pages: 1105

ISBN-13: 1603273751

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Recent advances in the biosciences have led to a range of powerful new technologies, particularly nucleic acid, protein and cell-based methodologies. The most recent insights have come to affect how scientists investigate and define cellular processes at the molecular level. This book expands upon the techniques included in the first edition, providing theory, outlines of practical procedures, and applications for a range of techniques. Written by a well-established panel of research scientists, the book provides an up-to-date collection of methods used regularly in the authors’ own research programs.

Differential Display
Differential Display

Author: Ron A. Leslie

Publisher: OUP Oxford

Published: 2000-05-04

Total Pages: 282

ISBN-13: 0191565946

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One of the most challenging tasks facing the modern biological research laboratory is to make sense of the enormous amount of data being generated by various genome projects currently underway, and especially the human genome project. Understanding the ways in which genes are differentially expressed in various tissues and cell types, throughout ontogenetic development and in pathological processes, will go a long way towards understanding the function of all these 'new' genes and their protein products. Differential Display explains in detail how to perform the technique of RT-PCR Differential Display in various kinds of experimental biological systems. It also examines this technique in the context of other methods of studying differential gene expression such as subtractive hybridisation and the use of high-density gene microarrays combined with hybridisation techniques and automatic image analysis.