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Basic Cell Culture Protocols
In this fully revised edition of an established classic, expert researchers and clinicians describe in step-by-step detail updated techniques for the isolation and growth of major primary cell types, such as kidney proximal tubule cells, hepatocytes, keratinocytes, and cardiomyocytes. The authors offer readily reproducible new methods for the differentiation of embryonic stem (ES) cells into various hematopoietic cell types, for fetal thymic organ culture, and for the isolation and culture of specialized cell types, such as mammary progenitor cells, skeletal muscle myofibers, mesenchymal cells, neural stem cells, hematopoietic cells, stromal cell lines, and endothelial cells. Additional chapters describe new techniques (leukocyte rolling, isolation of side-population cells, and scalable production of ES-derived cells) and detail quality control methods for cell lines (detection and elimination of mycoplasma, DNA fingerprinting, and cytogenetic analysis).
Forensic DNA Typing Protocols
A state-of-the-art collection of readily reproducible laboratory methods for DNA identity analysis, including Y chromosome haplotyping, mtDNA, and SNP typing. The book offers well-tested protocols for DNA quantification using real-time PCR on forensic samples and for the determination of the number of amelogenine gene copies. For forensic geneticists, there are readily reproducible methods for species identification, ancient DNA, and pharmacogenetics. Additional chapters address new applications in the forensic genetics lab, such a species identification or typing of CYP polymorphisms for the analysis of adverse to drugs.
Genomics, Proteomics, and Clinical Bacteriology
Gazing into crystal balls is beyond the expertise of most scientists. Yet, as we look further into the 21st century, one does not have to be Nostradamus to predict that the current genomics and proteomics "revolution" will have an immense impact on medical bacteriology. This impact is already being re- ized in many academic departments, and although encroachment on routine diagnostic bacteriology, particularly in the hospital setting, is likely to occur at a slower pace, it remains nonetheless inevitable. Therefore, it is important that no one working in bacteriology should find themselves distanced from these fundamental developments. The involvement of all clinical bacteriologists is essen...
Peptide Synthesis and Applications
Hands-on experts describe in step-by-step detail the key methodologies of contemporary peptide synthesis and illustrate their numerous applications. The techniques presented include protocols for chemical ligation, the synthesis of cyclic and phosphotyrosine-containing peptides, lipoamino acid- and sugar-conjugated peptides, and peptide purification and analyses. Additional chapters detail methodologies and instrumentation for high-throughput peptide synthesis, many different applications of peptides as novel research tools and biological probes, and the design and application of fluorescent substrate-based peptides that can be used to determine the selectivity and activity of peptidases. A practical guide to the identification of proteins using mass spectrometric analyses of peptide mixtures is also included.
RNA Silencing
A collection of readily reproducible methods for the design, preparation, and use of RNAs for silencing gene expression in cells and organisms. The techniques range widely and include methods addressing the biochemical aspects of the silencing machinery, RNA silencing in non-mammalian organisms, and the in vivo delivery of siRNAs and silencing vectors. There are also techniques for designing, preparing, and using RNAs to silence gene expression, for fine-tuning regulation by targeting specific isoforms of a given gene, and for the study and use of microRNAs. The protocols follow the successful Methods in Molecular BiologyTM series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
The Vagus-Immune Connection
This comprehensive guide delves into the intricate relationship between the vagus nerve and immune regulation, providing invaluable insights and practical strategies for understanding chronic illnesses like migraine, anxiety, BPD, cognitive dysfunction, stroke, hypertension, IBS, and more. Engineer, inventor, and vagus nerve expert J. P. Errico distills the latest scientific research and technological advancements to offer an easy-to-understand guide to bolstering your immune system and enhancing resilience against pathogens and diseases. With The Vagus-Immune Connection, you will: Gain a deep understanding of how the vagus nerve influences the immune system, from its role in controlling inflammation to its impact on autoimmune conditions; Discover practical techniques to effectively manage stress, a major contributor to immune dysregulation; Enhance immune resilience and disease prevention; Optimise cellular energy metabolism to improve neurodevelopment and cognitive function. Whethe
Oligonucleotide Synthesis
A collection of powerful new techniques for oligonucleotide synthesis and for the use of modified oligonucleotides in biotechnology. Among the protocol highlights are a novel two-step process that yields a high purity, less costly, DNA, the synthesis of phosphorothioates using new sulfur transfer agents, the synthesis of LNA, peptide conjugation methods to improve cellular delivery and cell-specific targeting, and triple helix formation. The applications include using molecular beacons to monitor the PCR amplification process, nuclease footprinting to study the sequence-selective binding of small molecules of DNA, nucleic acid libraries, and the use of small interference RNA (siRNA) as an inhibitor of gene expression.
Protein NMR Techniques
When I was asked to edit the second edition of Protein NMR Techniques, my first thought was that the time was ripe for a new edition. The past several years have seen a surge in the development of novel methods that are truly revolutionizing our ability to characterize biological macromolecules in terms of speed, accuracy, and size limitations. I was particularly excited at the prospect of making these techniques accessible to all NMR labs and for the opportunity to ask the experts to divulge their hints and tips and to write, practically, about the methods. I commissioned 19 chapters with wide scope for Protein NMR Techniques, and the volume has been organized with numerous themes in mind. ...
Epidermal Cells
A collection of cutting-edge methods to analyze and manipulate epidermal cellprecursors and mature epidermal cells. These protocols cover different methods and models for culturing epidermal cells, for enriching very early epidermal progenitors, and for studying epidermal cell commitment and differentiation both in vitro and in vivo. Topics of special interest include the derivation, characterization, and utility of epidermal stem cells, mature epidermal cells and their characterization, and applications in regenerative medicine. These readily reproducible techniques broaden our understanding of the biology of epidermal cells and of their utility in normal tissue homeostasis and regenerative medicine applications.
Cell Migration
A collection of classic, novel, and state-of-the-art methods for the study of cell migration in cultured cells, different model organisms, and specialized cells in normal development and disease. Highlights include basic assays that apply to all cell migration studies in vitro, assays in various model organisms, and assays for cancer cells, endothelial cells, and neurons both in vitro and in animal models. The protocols follow the successful Methods in Molecular BiologyTM series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
