Library Science

Pui-Yan Kwok, MD, PhD, has assembled a comprehensive collection of readily reproducible techniques for the difficult process of single nucleotide polymorphisms (SNP) discovery and genotyping. These cutting-edge protocols for mutation/SNP detection utilize denaturing high-performance liquid chromatography (dHPLC), single-strand conformation polymorphism (SSCP), conformation-sensitive gel electrophoresis (CSGE), chemical cleavage, and direct sequencing. Equally powerful and up-to-date methods are given for genotyping SNPs, including molecular beacons, the Taqman assay, single-base extension approaches, pyrosequencing, ligation, the Invader assay, and primer extension with mass spectrometry detection.

Authors: P-Y. Kwok

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A collection of cutting-edge techniques for using capillary electrophoresis (CE) to analyze complex carbohydrates. These readily reproducible protocols provide methods for sample preparation, analysis of mono- and oligosaccharides, glycoproteins, and glycoconjugates. A useful appendix describes the structures of the most commonly encountered carbohydrate residues and olgosaccharides from mammalian and bacterial origins. Each protocol contains detailed information on reagents, apparatus, notes, comments, and tips on procedures.

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Leading researchers in the biological, chemical, and physical investigation of superantigens describe in step-by-step detail their best experimental techniques to assess the physical characteristics and biological effects of superantigens. Their protocols range from those for investigating the interactions of superantigens with cellular receptors to those for the analysis of their immunological and biological effects, including methods for using BIOcore to determine binding kinetics and establishing various lymphocyte cell culture systems. There are also accounts of such methods as the RNase protection assay, cytokine ELISA, FACS analysis, and cytokine production at the single cell level. Ed. T. Krakauer

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Hands-on laboratory experts describe for the novice investigator a host of novel technologies and molecular techniques specifically designed to study cellular immunology and promote its gene therapy applications. Presented in step-by-step detail to ensure ready reproducibility, these protocols range from flow cytometric techniques to detect cytokines and growth factors in different specimens, to methods for generating and expanding dendritic cells and hematopoietic progenitors from different origins. Although the protocols require little previous experience, even highly skilled researchers will augment their skills with many novel timesaving techniques. Ed. D. Korholz and W. Kiess

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Hands-on laboratory experts present a set of “classic” PCR-based methods for the identification and detection of important animal and food microbial pathogens, including several zoonotic agents. These proven techniques can be precisely applied to a wide variety of microbes, among them Campylobacter spp., chlamydiae, toxigenic clostridia, Escherichia coli (STEC), Listeria monocytogenes, mycoplasmas, salmonellae, and Yersinia enterocolitica. Additional chapters review the specificity and performance of diagnostic PCR analysis, the pre-PCR processing of samples, the critical aspects of standardizing PCR methods, and the general issues involved in using PCR technology for microbial diagnosis.

Authors: Sachse, Konrad; Frey, Joachim Read the rest of this entry »

An international panel of recognized academic physicians, researchers, and clinical laboratory diagnosticians describe their best methods for characterizing neurologically relevant genes, their mutations, and their proteins. Providing detailed step-by-step instructions to assure successful experimental results, these experts cover the key methods for mutation detection and screening, including discussions of quantitative PCR, trinucleotide repeat detection, sequence-based mutation detection, fluorescence in situ hybridization (FISH), in vitro protein expression systems, and studies of protein expression and function. Ed. Potter, Nicholas T.

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A collection of readily reproducible experimental methods for studying the subversion of normal cell signaling that leads to cancer. Presented in step-by-step detail to ensure successful results, the methods include proven techniques for the investigation of apoptosis and cell death, complementary protocols for manipulating and/or monitoring oncogenic signals in cancer cells, and techniques for studying protein–protein interactions. A detailed protocol for capturing pure samples of malignant cells from frozen tissue specimens and two alternative techniques for analyzing their genomic DNA is also described.

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A panel of leading scientific experts detail novel techniques and strategies for the cellular and genetic modification of heart function. The highly experienced authors provide step-by-step protocols for vector production and purification, for gene and cell delivery techniques, and for physiological assessment in vivo and in vitro. Timely, authoritative, and state-of-the-art, Cardiac Cell and Gene Transfer: Principles, Protocols, and Applications constitutes an invaluable guide to all the new cellular and gene-based technologies needed by basic and clinical investigators working to illuminate today’s unanswered questions about heart disease and ultimately to improve the heart performance in all their patients.

Ed. Metzger, Joseph M.

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A collection of key cytogenetic and FISH techniques used by modern clinical laboratories in the genetic analysis of human malignancies. The book’s practical advice and methods are suitable for use at every level of expertise, including fully established laboratories, but with a sympathetic bias towards anyone considering setting up a new cytogenetics service. Here the reader will find not only elementary tutorials on the fundamentals of human karyotypes and chromosome analysis, but also detailed discussions on how laboratories may optimally upgrade their repertoire of capabilities to include such newer complementary techniques as CGH, FISH, and M-FISH.  Ed. Swansbury, John

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Expert researchers and inventors in the field describe their own proven techniques for generating cDNA/mRNA libraries to identify the functions of specific decoded gene sequences. A wide variety of techniques is presented for enhancing the generation of complete and full-length libraries, and for confirming the quality of the cDNAs generated. Among the applications detailed are electrophoresis, Northern blotting, single cell microarray analysis, subtractive hybridization, subtractive cloning, gene cloning, and peptide library generation.  Ed. S.-Y. Ying

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Leading physician scientists and academic researchers review all the known tumor suppressor genes, explain how they work, and describe how they were discovered and isolated. In many cases, the authors discuss specific genes that are frequently involved in hereditary or sporadic cancers. They also provide a detailed guide to using powerful molecular genetic, cytogenetic, proteomic, and cell biological strategies to discover and isolate novel tumor suppressor genes and their targets. For both volumes: Leading physician scientists and academic researchers review all the known tumor suppressor genes, explain how they work, and describe how they were discovered and isolated. In many cases, the authors discuss specific genes that are frequently involved in hereditary or sporadic cancers. They also provide a detailed guide to using powerful molecular genetic, cytogenetic, proteomic, and cell biological strategies to discover and isolate novel tumor suppressor genes and their targets. A second volume of this two-volume set, Tumor Suppressor Genes, Volume 2: Regulation, Function, and Medical Applications, shows how to explore the cell biology and biochemical function of such encoded proteins, to study its physiological role in vivo, and to use information on TSGs to develop diagnostic and therapeutic strategies for cancer.      Ed. El-Deiry, Wafik S.

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