Molecular Microbial Ecology Manual

For a long time microbial ecology has been developed as a distinct field within Ecology. In spite of the important role of microorganisms in the environment, this group of 'invisible' organisms remained unaccessable to other ecologists. Detection and identification of microorganisms remain largely dependent on isolation techniques and characterisation of pure cul tures. We now realise that only a minor fraction of the microbial com munity can be cultivated. As a result of the introduction of molecular methods, microbes can now be detected and identified at the DNA/RNA level in their natural environment. This has opened a new field in ecology: Molecular Microbial Ecology. In the present manual we aim to introduce the microbial ecologist to a selected number of current molecular techniques that are relevant in micro bial ecology. The first edition of the manual contains 33 chapters and an equal number of additional chapters will be added this year. Since the field of molecular ecology is in a continuous progress, we aim to update and extend the Manual regularly and will invite anyone to depo sit their new protocols in full detail in the next edition of this Manual. We hope this book finds its place where it was born: at the lab bench! Antoon D.L. Akkermans, Jan Dirk van Elsas and Frans J. de Bruijn March 1995 Molecular Microbial Ecology Manual 1.3.6: 1-8, 1996. © 1996 Kluwer Academic Publishers.

Microbes are key drivers of the world's ecosystems. The vast majority of the world's diversity and metabolic potential lies within micro-organisms, yet we are just beginning to understand and utilize this ultimate resource of biological diversity. Critical to our exploration of the microbial world are methods that allow for the analysis of organisms that are invisible to our eyes, difficult to distinguish from each other, and often impossible to grow using available culture methods. The field of microbial ecology has been revolutionized in the past two decades by the introduction of molecular methods into the toolbox of the microbial ecologist. This molecular arsenal has helped to unveil the enormity of microbial diversity across the breadth of the earth's ecosystems, and has revealed that we are only familiar with a very small minority of the organisms that carry out key microbial functions in diverse habitats. The Molecular Microbial Ecology Manual, Second Edition (MMEM-II) provides a detailed and user-friendly description of the methods that have made this revolution in microbial ecology possible. However, what is perhaps most exciting about MMEM-II is that it contains a large number of new chapters, highlighting the newest trends in microbial ecology research, which seek to provide more quantitative and statistically robust data, and means of coupling microbial identity and function. In addition, the majority of the proven methods described in MMEM's first version have undergone significant revisions to provide the most up-to-date applications available. The state-of-the-art methods described in MMEM-II have not only been provided by experts in the field, but in most cases by the laboratories that actually first developed and applied the methods, thus providing the MMEM-II user with unique first-hand tips and insight. The new on-line format available forMMEM-II should also add to the utility of MMEM-II by allowing users to search for key topics throughout the manual, skip between interrelated chapters at the push of a button, and by providing immediate availability to protocol updates and new chapters dedicated to future technical developments.

The premiere two-volume reference on revelations from studying complex microbial communities in many distinct habitats Metagenomics is an emerging field that has changed the way microbiologists study microorganisms. It involves the genomic analysis of microorganisms by extraction and cloning of DNA from a group of microorganisms, or the direct use of the purified DNA or RNA for sequencing, which allows scientists to bypass the usual protocol of isolating and culturing individual microbial species. This method is now used in laboratories across the globe to study microorganism diversity and for isolating novel medical and industrial compounds. Handbook of Molecular Microbial Ecology is the first comprehensive two-volume reference to cover unculturable microorganisms in a large variety of habitats, which could not previously have been analyzed without metagenomic methodology. It features review articles as well as a large number of case studies, based largely on original publications and written by international experts. This first volume, Metagenomics and Complementary Approaches, covers such topics as: Background information on DNA reassociation and use of 16 rRNA and other DNA fingerprinting approaches Species designation in microbiology Metagenomics: Introduction to the basic tools with examples Consortia and databases Bioinformatics Computer-assisted analysis Complementary approaches—microarrays, metatranscriptomics, metaproteomics, metabolomics, and single cell analysis A special feature of this volume is the highlighting of the databases and computer programs used in each study; they are listed along with their sites in order to facilitate the computer-assisted analysis of the vast amount of data generated by metagenomic studies. Handbook of Molecular Microbial Ecology I is an invaluable reference for researchers in metagenomics, microbiology, and environmental microbiology; those working on the Human Microbiome Project; microbial geneticists; molecular microbial ecologists; and professionals in molecular microbiology and bioinformatics.

The premiere two-volume reference on revelations from studying complex microbial communities in many distinct habitats Metagenomics is an emerging field that has changed the way microbiologists study microorganisms. It involves the genomic analysis of microorganisms by extraction and cloning of DNA from a group of microorganisms, or the direct use of the purified DNA or RNA for sequencing, which allows scientists to bypass the usual protocol of isolating and culturing individual microbial species. This method is now used in laboratories across the globe to study microorganism diversity and for isolating novel medical and industrial compounds. Handbook of Molecular Microbial Ecology is the first comprehensive two-volume reference to cover unculturable microorganisms in a large variety of habitats, which could not previously have been analyzed without metagenomic methodology. It features review articles as well as a large number of case studies, based largely on original publications and written by international experts. This second volume, Metagenomics in Different Habitats, covers such topics as: Viral genomes Metagenomics studies in a variety of habitats, including marine environments and lakes, soil, and human and animal digestive tracts Other habitats, including those involving microbiome diversity in human saliva and functional intestinal metagenomics; diversity of archaea in terrestrial hot springs; and microbial communities living at the surface of building stones Biodegradation Biocatalysts and natural products A special feature of this book is the highlighting of the databases and computer programs used in each study; they are listed along with their sites in order to facilitate the computer-assisted analysis of the vast amount of data generated by metagenomic studies. Such studies in a variety of habitats are described here, which present a large number of different system-dependent approaches in greatly differing habitats. Handbook of Molecular Microbial Ecology II is an invaluable reference for researchers in metagenomics, microbial ecology, microbiology, and environmental microbiology; those working on the Human Microbiome Project; microbial geneticists; and professionals in molecular microbiology and bioinformatics.

Section 1: Isolation of nucleic acids. Section 2: Detection of microbial nucleic acid sequences. Section 3: Identification and classification of microbes using DNA and RNA sequences. Section 4: Detection, identification and classification of microbes using other methods. Section 5: Detection of gene transfer in the environment. Section 6: Tracking of specific microbes in the environment. Section 7: Statistical, computer-assisted and other analyses. Section 8: Molecular tools to assess microbial activities.

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.