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Recombinant DNA Principles and Methodologies
This comprehensive yet balanced work emphasizes the principles and rationale underlying recombinant DNA methodology while furnishing a general understanding of the experimental protocols-suggesting flexible approaches to resolving particular molecular necessities that are easily adaptable to readers' specific applications. Features summary tables presenting at-a-glance information on practices of recombinant DNA methodologies! Recombinant DNA Principles and Methodologies discusses basic and advanced topics requisite to the employment of recombinant DNA technology, such as plasmid biology nucleic acid biochemistry restriction enzymes cloning strategies gel electrophoresis southern and norther...
Trinucleotide Repeat Protocols
Trinucleotide repeats are relatively common in the human genome. These simple repeats have received much attention since epoch-making discoveries were made that particular trinucleotide repeats are expanded in the causal genes of human hereditary neurological disorders. For example, the CGG repeat is expanded in fragile X syndrome at the 5' untranslated region (UTR) of its causal gene. In myotonic dystrophy, it is the CTG repeat that is expanded at the 3' UTR of its causal gene. The CAG repeat was also found expanded in coding regions of the genes responsible for X-linked spinal and bulbar muscular atrophy, Huntington’s disease, spinocerebellar ataxia, and other disorders. On the other han...
Capillary Electrophoresis of Proteins and Peptides
Throughout the more than 20 years that have followed the beginnings of capillary electrophoresis (CE), its application to the analysis of proteins and peptides has continued to be reliable, versatile, and productive. Over time, CE has matured to become a superb complement to HLPC, and in many cases has also evolved as an automated and quantitative replacement for conventional slab gel electrophoresis methods such as SDS-PAGE and isoelectric focusing. Within Capillary Electrophoresis of Proteins and Peptides, we have assembled contributions from researchers who are applying state-of-the-art CE for protein and peptide analysis, including topics that we believe are of great potential both in th...
Epigenetics Protocols
The field of epigenetics has grown exponentially in the past decade, and a steady flow of exciting discoveries in this area has served to move it to the forefront of molecular biology. Although epigenetics may previously have been considered a peripheral science, recent advances have shown considerable progress in unraveling the many mysteries of nontraditional genetic processes. Given the fast pace of epigenetic discoveries and the groundbreaking nature of these developments, a thorough treatment of the methods in the area seems timely and appropriate and is the goal of Epigenetics Protocols. The scope of epigenetics is vast, and an exhaustive analysis of all of the techniques employed by i...
Nitric Oxide Protocols
Carrying on the high standards of the much-praised first edition of Nitric Oxide Protocols, Aviv Hassid has brought together a panel of expert researchers and clinician scientists to describe in step-by-step detail the latest methodologies for the measurement of nitric oxide--and the enzyme that produces it-in biological tissues and fluids. The authors take advantage of the latest methodologies for the quantitation of biological fluids and tissues, including capillary electrophoresis, microcoaxial electrodes, in vivo measurement of nitric oxide in exhaled air, confocal microscopy, gas chromatography, in situ hybridization, and real-time polymerase chain reaction. Chapters on the measurement ...
Checkpoint Controls and Cancer
Intracellular checkpoint controls constitute a network of signal transd- tion pathways that protect cells from external stresses and internal errors. Ext- nal stresses can be generated by the continuous assault of DNA-damaging agents, such as environmental mutagens, ultraviolet (UV) light, ionizing radiation, or the reactive oxygen species that can arise during normal cellular metabolism. In response to any of these assaults on the integrity of the genome, the activation of the network of checkpoint control pathways can lead to diverse cellular responses, such as cell cycle arrest, DNA repair, or elimination of the cell by cell death (apoptosis) if the damage cannot be repaired. Moreover, in...
Apoptosis Methods and Protocols
The most fundamental question facing each and every cell within an org- ism is to survive or to die. Cell death is required for normal function; some estimates suggest that as many as one million cells undergo cell death every second in the adult human body. Almost all cells undergoing physiological, or programmed, cell death, independent of cell type, manifest a stereotypic p- tern of morphological changes termed apoptosis. Typically, apoptotic cells d- play shrinkage, membrane blebbing, chromatin condensation, and nuclear fragmentation. The integrity of the cell membrane is not lost during apoptosis and so avoids eliciting the inflammatory response that would have been caused by the spilla...
Ribozymes and siRNA protocols
In this completely updated and expanded edition of a classic bench manual, hands-on experts take advantage of the latest advances in ribozyme, DNAzyme, hammerhead ribozymes and derivatives, and RNA interference technologies to describe in detail the exciting and successful methods now available for gene inactivation in vitro and in vivo. Their optimized techniques employ hairpin ribozymes, DNAzymes, hammerhead ribozymes and derivatives, group I intron ribozymes, RNase P ribozymes, and siRNAs, as well as general methods for RNA structure analysis, delivery of oligonucleotides, and gene therapy. Also provided are novel methods for identifying accessible cellular mRNA sites; group I intron and ...
Platelets and Megakaryocytes
12 The average human body has in the order of 10 circulating platelets. They are crucial for hemostasis, and yet excessive platelet activation is a major cause of m- bidity and mortality in western societies. It is therefore not surprising that platelets have become one of the most extensively investigated biological cell types. We are, however, far from understanding precisely how platelets become activated under physiological and pathophysiological conditions. In addition, there are large gaps in our knowledge of platelet production from their giant precursor cell, the megakar- cyte. Understanding megakaryocyte biology will be crucial for the development of platelet gene targeting. The aim...
Cell Cycle Control and Dysregulation Protocols
Cell Cycle Control and Dysregulation Protocols focuses on emerging methodologies for studying the cell cycle, kinases, and kinase inhibitors. It addresses the issue of gene expression in vivo and in vitro, the analysis of cyclin-dependent kinase inhibitors, protein degradation mediated by the proteosome, the analysis of the transformed cell phenotype, and innovative techniques to detect apoptosis. Because there are already many manuals and protocols available, along with commercial kits and reagents, a variety of the more common techniques have not been included in our book. The protocols described, based on rather sophisticated techniques for in vivo and in vitro studies, consist of molecular biology, biochemistry, and various types of immunoassays. Indeed, the authors have successfully accomplished an arduous task by presenting several topics in the simplest possible manner. We are confident that Cell Cycle Control and Dysregulation Protocols will facilitate and optimize the work of practical scientists involved in researching the cell cycle. We greatly acknowledge the extraordinary contribution of the authors in writing this book.
