Advanced Methods – serie

DNA Microarrays introduces all up-to-date microarray platforms and their various applications. It is written for scientists who are entering the field of DNA microarrays as well as those already familiar with the technology, but interested in new applications and methods.

With a variety of detection chemistries, an increasing number of platforms, multiple choices for analytical methods and the jargon emerging along with these developments, real-time PCR is facing the risk of becoming an intimidating method, especially for beginners. Real-time PCR provides the basics, explains how they are exploited to run a real-time PCR assay, how the assays are run and where these assays are informative in real life. It addresses the most practical aspects of the techniques with the emphasis on 'how to do it in the laboratory'. Keeping with the spirit of the Advanced Methods Series, most chapters provide an experimental protocol as an example of a specific assay.

Microoganisms are distributed across every ecosystem, and microbial transformations are fundamental to the operation of the biosphere. Microbial ecology is the study of this interaction between microorganisms and their environment, and arguably represents one of the most important areas of biological research. Yet for many years our study of microbial flora was severely limited: the primary method of culturing microorganisms on media allowed us to study only between 0.1 and 10% of the total microbial flora in any given environment.Molecular Microbial Ecology gives a comprehensive guide to the recent revolution in the study of microorganisms in the environment. Details are given on molecular methods for isolating some of the previously uncultured and numerically dominant microbial groups. PCR-based approaches to studying prokaryotic systematics are described, including ribosomal RNA analysis and stable isotope probing. Later chapters cover DNA hybridisation techniques (including fluorescent in situ hybridisation), as well as genomic and metagenomic approaches to microbial ecology. Gathering together some of the world’s leading experts, this book provides an invaluable introduction to the modern theory and molecular methods used in studying microbial ecology.

Gene Targeting and Embryonic Stem Cells is a practical guide designed for the rapidly growing number of researchers who are moving into this field. Provides details on how to culture, transfect and differentiate established cell lines, and how to isolate new cell lines. Gene targeting experiments are described for a number of cell types, including ungulate fetal fibroblasts, murine ES cells, human embryonal carinoma cells and human ES cells, and include protocols for gene-targeting vectors, DNA transfection and RNA interference. The recent isolation of human embryonic stem cells and the potential of these cells for therapeutic applications has generated an entirely new methodology. Similarly, gene targeting methodology has recently been extended to nuclear donor cells in ungulate species. This volume will be invaluable for both new and established researchers in the field of human embryonic stem cells, and to biotech companies engaged in the production of transgenic proteins in livestock, xenotransplantation and the development of animal models.

One of the major recent discoveries in molecular and cellular biology is that small double-stranded RNA molecules selectively turn off gene expression in all types of cell, a phenomenon known as RNA silencing. This discovery led to the development of RNA interference (known as RNAi) as a powerful research tool in the functional study of individual genes and their products, and in functional genomics. In RNAi, specific small double-stranded RNA molecules (small interfering RNAs or siRNAs) are introduced into cells to selectively silence certain genes.RNAi covers the basic concepts and mechanisms of RNAi, transfection of cells with siRNAs, the design and validation of RNAi reagents, RNAi techniques in different organisms, large-scale RNAi screening, applications of RNAi in drug discovery, and potential uses of RNAi as a therapeutic agent. A key feature of RNAi is the highlighting of the pitfalls that can occur and how to minimize them. The book also contains a complete list of abbreviations.

The 'Advanced Methods' series is intended for advanced undergraduates, postgraduates and established research scientists. Titles in the series are designed to cover current important areas of research in life sciences, and include both theoretical background and detailed protocols. The aim is to give researchers sufficient theory, supported by references, to take the given protocols and adapt them to their particular experimental systems.Microarrays and Microplates title looks at the new microarray and microplate-based technologies which facilitate large-scale analysis of DNA sequence variants, mRNA molecules and proteins. The book provides a review of the various methodologies being used to identify genetic variants and gene regulation and guides readers through both the application of these methodologies and experimental procedures.