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 »

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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Prominent experts from around the world detail the chromatographic and electroseparation techniques they have developed for chiral separations on an analytical scale. Described in step-by-step detail to ensure successful experimental results, the procedures are presented as either general methods or as specific applications to substance classes and special compounds, with emphasis on high performance liquid chromatography and capillary electrophoresis techniques, but also including thin layer chromotographic, gas chromatographic, supercritical fluid chromatographic as well as recent electrochromotographic techniques.

Ed. Gbitz, Gerald; Schmid, Martin G.

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The first compilation of classic and emerging laboratory techniques for the study of the biochemistry and cell biology of protein kinase C (PKC). Described in step-by-step detail, these methods can be easily used to explore the structure, function, regulation, subcellular localization, and macromolecular interactions of protein kinase C.. Each protocol is introduced in the context of PKC function and regulation and contains many notes on how best to deal with the problems that may occur. Comprehensive and authoritative, Protein Kinase C Protocols is a timely compilation of biophysical, biochemical, cell biological, and molecular biological approaches that brings protein kinase C research into any laboratory interested in studying it.

Ed. Newton, Alexandra C. ISBN: 1588290689

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Book Description

An indispensable collection of novel techniques that have proven most useful for studying the physiological properties of p53 both in vitro and in vivo. The techniques provide proven solutions to problems in studying the purification, target identification, gene expression, quantitation, interaction, signaling, transactivation, and transrepression of p53. The methods are also useful for delineating the functions of other proteins that may act as tumor or growth suppressors. Each technique includes step-by-step instructions, troubleshooting notes, a theoretical review, and discussion of associated problems that might arise during the course of investigation.

Authors: Deb, Sumitra; Deb, Swati Palit

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Authors:Casali, Nicola; Preston, Andrew

A comprehensive collection of readily reproducible techniques for the manipulation of recombinant plasmids using the bacterial host E. coli. The authors describe proven methods for cloning DNA into plasmid vectors, transforming plasmids into E. coli, and analyzing recombinant clones. They also include protocols for the construction and screening of libraries, as well as specific techniques for specialized cloning vehicles, such as cosmids, bacterial artificial chromosomes, l vectors, and phagemids. Common downstream applications such as mutagenesis of plasmids, recombinant protein expression, and the use of reporter genes, are also described.

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Erich Grotewold has assembled a team of leading plant scientists to describe in detail the most commonly used methods for investigating plant gene function in a wide variety of plants, during plant pathogen interactions, and even in algae. These readily reproducible protocols include computational, molecular, and genetic methodologies designed for both general and specific problems. Here the reader will learn about powerful computational and statistical tools to help predict gene function either on the basis of comparative genomics, or from the analysis of complex genome sequences. Numerous loss-of-function and gain-of-function techniques for discovering gene function are presented in step-by-step detail.

Authors: Grotewold, Erich

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