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 This may come as a surprise to many, but to us it implies that many of the DNA microarray studies went unpublished owing to their unfulfilled promises (1).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.


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.


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.


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


Aging Methods and Protocols

Aging Methods and Protocols

Author: Yvonne A. Barnett

Publisher: Springer Science & Business Media

Published: 2008-02-01

Total Pages: 378

ISBN-13: 1592590705

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Aging is an almost universal process within biological systems, one which leads to a decline in functional capacity, disease onset, and eventually death. There has been much interest in recent years to elucidate the molecular mec- nisms that underlie the aging process. Many theories have been proposed since the last century that aim to explain the causes of aging. There is no one theory that completely satisfies the phenotype of aging, but genetics and environm- tal factors play an important role in the etiology of age-related pathologies and the aging process. However, there is still much to be learned about the aging process which has been termed one of the last great frontiers in biology. De- graphic changes worldwide are leading to increased average life expectancies within our populations. These changes in population characteristics will impact upon the economies of the supporting society, with increasing healthcare and infrastructural costs arising from the prevalence of age-related pathologies and other physical disabilities associated with advancing years. Many researchers worldwide are working in the attempt to identify key cellular processes through which it might one day be possible to slow down the aging process and thus increase the health span of humans. Numerous research projects—from the cellular through to tissue, organ, and whole organism studies—are currently underway to investigate the mul- factorial aging process.


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.


Handbook of Molecular-Genetic Techniques for Brain and Behavior Research

Handbook of Molecular-Genetic Techniques for Brain and Behavior Research

Author: Wim E. Crusio

Publisher: Elsevier

Published: 1999-10-18

Total Pages: 994

ISBN-13: 0080536867

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The book gives a broad overview of recombinant DNA techniques for the behavioral neuroscientist, with illustrative examples of applications. Species covered include rodents (mainly mice), Drosophila melanogaster, Caenorhabditis elegans and Danio rerio. Experimental techniques required to characterize the behavioral phenotypes of mutant animals is provided. Several aspects of novel molecular-genetic techniques are overviewed and possible research strategies are explained. The sections of the book start with general descriptions of techniques followed by illustrative examples. It is divided into six sections. Section 1, bioinformatics and genomics research. Section 2, top-down strategies, where the researcher starts with the phenotype and then analyzes the associated genes; bottom-up strategies, where the physiological chain leading to a phenotype is analyzed starting from the gene product. Section 3, transgenic approaches in rodents including overexpressing foreign genes and gene-targeting; systemic manipulation approaches directly targeting the central nervous system and methods used with invertebrates. Section 4, methods used to evaluate relevant behavioral phenotypes, including learning and aggression. Section 5, examples on molecular brain research in man. Section 6, ethical aspects of research in this field.


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 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.