Bøger i Springer Protocols Handbooks serien

The book provides the readers of various discipline an easy understanding of the latest biophysical techniques pertaining to microbiology. Biofilm associated chronic infection is a major health problem and a serious concern to doctors, scientists and other health workers as it develops antibiotic and multi-drug resistance. This book describes various protocols utilized in the detection of the biofilm. The book has been divided into six sub sections which provides pertinent information about the various biophysical techniques and instruments that are used for detecting and analyzing the biofilm formation upon biotic and abiotic surfaces. The readers will be able to identify the techniques that can best cater information to solve the problem at hand. This book attempts to compile the latest information on the recent advances in the various functional aspects of microbial biofilms, their pathogenesis, present day treatments as well as detection strategies.This book is meant forresearchers in the field of microbiology and interested in understanding microbial pathogenesis, quorum sensing and biofilm formation.

This volume details methods and protocols necessary to further the study of insect immunity. Chapters guide readers through up-to-date genomic and transcriptomic approaches, insect samples for proteomic analysis, hemocytes in Drosophila, cellular response in Lepidoptera, insect AMPs, manipulate the composition of mosquito microbiota, viral infections in insects, infections by entomopathogenic nematodes, immune response following oral infections, and protocols to to monitor the effect of septic infections with human pathogens using B. mori as a model. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Immunity in Insects aims to ensure successful resultsin the further study of this vital field.

A comprehensive treasury of the key molecular biology methods - ranging from DNA extraction to gene localization in situ - needed to function effectively in the modern laboratory. Each of the 120 highly successful techniques follows the format of the much acclaimed "Methods in Molecular Biology" series.

This Handbook reviews neuroprotection based on contemporary understanding of the molecular basis of neurological disorders. Cutting-edge and innovative, the book documents neuroprotective effects of established drugs, as well as new drugs in development.

There are also readily reproducible methods for protein expression profiling, identifying protein-protein interactions, and protein chip technology, as well as a range of newly developed methodologies for determining the structure and function of a protein.

This Volume presents applications of hydrocarbon microbiology in the context of environmental pollutant degradation, covering pollutants such as petroleum and related wastes (i.e.

This Volume presents methods for quantifying microbial populations and characterising microbial communities by extracting and analysing biomarkers such as RNA, DNA and lipids. The chapters cover a wide range of topics, including: cell separation from oil-rich environments, enumeration of hydrocarbon degraders and sulphate reducers using most-probable-number techniques, and quantification by means of real-time PCR. A variety of molecular methods are described for microbial community profiling, such as phospholipid fatty acid analysis, DGGE, T-RFLP and SSCP. One chapter examines high-throughput sequencing, and provides important information on the associated procedures required for thorough data analysis. A further chapter is devoted to the characterisation of protistan communities, while the closing chapter describes multiplex fluorescent antibody microarrays for detecting microbial biomarkers.Hydrocarbon and Lipid Microbiology ProtocolsThere are tens of thousands of structurally different hydrocarbons, hydrocarbon derivatives and lipids, and a wide array of these molecules are required for cells to function. The global hydrocarbon cycle, which is largely driven by microorganisms, has a major impact on our environment and climate. Microbes are responsible for cleaning up the environmental pollution caused by the exploitation of hydrocarbon reservoirs and will also be pivotal in reducing our reliance on fossil fuels by providing biofuels, plastics and industrial chemicals. Gaining an understanding of the relevant functions of the wide range of microbes that produce, consume and modify hydrocarbons and related compounds will be key to responding to these challenges. This comprehensive collection of current and emerging protocols will facilitate acquisition of this understanding and exploitation of useful activities of such microbes.

This manual offers detailed protocols for fluorescence in situ hybridization (FISH) and comparative genomic hybridization approaches, which have been successfully used to study various aspects of genomic behavior and alterations.

This Volume presents relevant single-cell and single-molecule approaches in the study of microbes producing and utilizing hydrocarbons and lipids. While generically applicable for all microorganisms, the approaches described are, wherever possible, adapted to the field of study of hydrocarbon and lipid microbiology. The methods include basic procedures for isolating single cells by means of microfluidics and flow cytometry, and their cultivation in arrays as pure clones; for isolating, amplifying and sequencing single-cell genomes and transcriptomes; and for analysing single-cell metabolomes by means of Raman spectroscopy. Single-molecule approaches include the use of protein:fluorescent dye fusions for protein localization and methods for the production of cell division protostructures and lipid monolayers. Methods for the functional analysis of single cells include detection of metabolically active (protein-synthesizing) cells in environmental samples by bioorthogonal non-canonical amino acid tagging, Raman spectroscopy combined with stable isotope labelling and fluorescent in situ hybridisation, and visualization of single cells participating in gene transfer activity. Lastly, protocols are presented for single-cell biotechnological applications, including biofuel production.Hydrocarbon and Lipid Microbiology ProtocolsThere are tens of thousands of structurally different hydrocarbons, hydrocarbon derivatives and lipids, and a wide array of these molecules are required for cells to function. The global hydrocarbon cycle, which is largely driven by microorganisms, has a major impact on our environment and climate. Microbes are responsible for cleaning up the environmental pollution caused by the exploitation of hydrocarbon reservoirs and will also be pivotal in reducing our reliance on fossil fuels by providing biofuels, plastics and industrial chemicals. Gaining an understanding of the relevant functions of the wide range of microbes that produce, consume and modify hydrocarbons and related compounds will be key to responding to these challenges. This comprehensive collection of current and emerging protocols will facilitate acquisition of this understanding and exploitation of useful activities of such microbes.

This Volume presents key microscopy and imaging methods for revealing the structure and ultrastructure of environmental and experimental samples, of microbial communities and cultures, and of individual cells.

< Hydrocarbon and Lipid Microbiology ProtocolsThere are tens of thousands of structurally different hydrocarbons, hydrocarbon derivatives and lipids, and a wide array of these molecules are required for cells to function.

This Volume presents a comprehensive series of generic protocols for the genetic and genomic analysis of prokaryotic isolates. Genetic methods for functional analyses employ the latest cloning vectors, gene fusion methods and transposon mutagenesis systems, as well as systems for introducing protease-cleavage sequences into permissive sites in proteins under investigation. Genomic methods described include protocols for transcriptomics, shotgun proteomics, interactomics, metabolic profiling, and lipidomics. Bioinformatic tools for genome annotation, transcriptome display and the integration of transcriptomic data into genome-scale metabolic reconstructions are described. Protocols for 13C-based metabolic flux determinations and analysis of the hierarchical and metabolic regulation of fluxes through pathways are included. The Volume thus enables investigators to functionally analyse an isolate over the entire cellular range spanning the gene, the genome, the transcript repertoire, the proteome, the interactome, the metabolic network with its nodes and their regulatory hierarchies, and the metabolic fluxes and their physiological controls.Hydrocarbon and Lipid Microbiology ProtocolsThere are tens of thousands of structurally different hydrocarbons, hydrocarbon derivatives and lipids, and a wide array of these molecules are required for cells to function. The global hydrocarbon cycle, which is largely driven by microorganisms, has a major impact on our environment and climate. Microbes are responsible for cleaning up the environmental pollution caused by the exploitation of hydrocarbon reservoirs and will also be pivotal in reducing our reliance on fossil fuels by providing biofuels, plastics and industrial chemicals. Gaining an understanding of the relevant functions of the wide range of microbes that produce, consume and modify hydrocarbons and related compounds will be key to responding to these challenges. This comprehensive collection of current and emerging protocols will facilitate acquisition of this understanding and exploitation of useful activities of such microbes.

This Volume covers protocols for various applications in hydrocarbon microbiology, including those of interest for industrial processes, biocatalysis, lipid and biofuel production, bioproducts, or the human microbiome.

Hydrocarbon and Lipid Microbiology ProtocolsThere are tens of thousands of structurally different hydrocarbons, hydrocarbon derivatives and lipids, and a wide array of these molecules are required for cells to function.

This Volume presents protocols for investigating the genetic, metabolic and ecological potential and functional analysis of microbial communities.

This practical volume brings together a broad range of topics related to membrane biology research with particular emphasis on novel approaches, technology platforms, and emerging tools in this area of study.

It explores conventional immunohistochemistry, virus neutralization, enzyme-linked immunosorbent assays, expression and purification of recombinant viral proteins, and various molecular assays, including conventional and real-time reverse transcription-PCR, reverse genetics methodology, and next generation sequencing and sequence analyses.

This Volume features protocols for investigating the hydrocarbon- and lipid-specific activities of microbes. They include methods for studying chemotaxis, the colonisation of hydrocarbon surfaces, hydrocarbon uptake, respiration, nitrogen fixation, sulphate reduction, membrane stabilisation through cis-trans isomerisation of membrane fatty acids, and the production of biosurfactants and biopolymers in response to the presence of hydrocarbons. A protocol for studying the ability of microbes to control the concentration of hydrocarbons in their aqueous environment is also described, and phenotyping methods to reveal microbes' more general metabolic activities are presented. Several protocols for investigating acid production in connection with oil souring and biocorrosion by microbes in oil well, oil transportation and storage settings are presented. Lastly, protocols for measuring methanogenesis, as an example of microbial hydrocarbon production, are described.

The aim of this book is to improve pyrosequencing protocols as well as instrumentation for better clinical use by describing improvements and novel applications of pyrosequencing technology.

This volume includes in-depth, hands-on protocols for detecting various developmental defects in Drosophila.

Since the successes in a laboratory setting do not guarantee plant survival and propagation in greenhouses and in the natural environment, it discusses greenhouse propagation techniques that are essential to the survival of plants generated from a laboratory setting.

This protocol book includes common and advanced research protocols for Yersinia pestis research and for its identification, genotyping and source-tracing. This book includes protocols for genomic and transcriptomic analysis, small RNA study, protein-protein interaction, gene deletion.

This new volume, besides bringing new and original aspects of what is found in the literature, fills some of the gaps in volume one by including the most systematic and extensive treatment of methods and techniques in qualitative research.

This book provides practical information on a whole set of protein experiments for advanced structural biology, such as X-ray crystallography, NMR, electron microscopy, advanced mass spectroscopy, and surface plasmon resonance, as well as a wide variety of expression systems including eukaryotic and in vitro expression.

This Volume describes methods for investigating microbes in their natural environment and how to obtain representative samples and preserve them for subsequent analyses. Chapters are arranged according to the environments under investigation, which include: oil reservoirs, fracking fluids, aquifers, coal beds, oil sands and their tailing ponds, lakes, rivers, leaves, polar seas and ice, the sea-surface microlayer, mud flats, microbialites, and deep-sea fauna. A variety of downstream analytical procedures are described, including: nucleic-acid extraction and preparation for high-throughput sequencing, fluorescence in-situ hybridisation, and cultivation of aerobic and anaerobic hydrocarbon-degrading microbes. Though most chapters focus on hydrocarbon-rich environments, many of the approaches used are generic, and as such will be of value to researchers embarking on studies of microbes and their processes in the field.Hydrocarbon and Lipid Microbiology Protocols There are tens of thousands of structurally different hydrocarbons, hydrocarbon derivatives and lipids, and a wide array of these molecules are required for cells to function. The global hydrocarbon cycle, which is largely driven by microorganisms, has a major impact on our environment and climate. Microbes are responsible for cleaning up the environmental pollution caused by the exploitation of hydrocarbon reservoirs and will also be pivotal in reducing our reliance on fossil fuels by providing biofuels, plastics and industrial chemicals. Gaining an understanding of the relevant functions of the wide range of microbes that produce, consume and modify hydrocarbons and related compounds will be key to responding to these challenges. This comprehensive collection of current and emerging protocols will facilitate acquisition of this understanding and exploitation of useful activities of such microbes.

This Volume addresses the pros and cons of oligonucleotide probes, primers and primer combinations, and importantly considers how to design the best tools for the microbial taxa and/or processes being investigated. Individual chapters focus on the design of primers targeting genes that code for enzymes associated with the following functions: degradation of aromatic, aliphatic and chlorinated hydrocarbons under aerobic and anaerobic conditions, methanogenesis, methane oxidation, and the nitrogen cycle.Hydrocarbon and Lipid Microbiology ProtocolsThere are tens of thousands of structurally different hydrocarbons, hydrocarbon derivatives and lipids, and a wide array of these molecules are required for cells to function. The global hydrocarbon cycle, which is largely driven by microorganisms, has a major impact on our environment and climate. Microbes are responsible for cleaning up the environmental pollution caused by the exploitation of hydrocarbon reservoirs and will also be pivotal in reducing our reliance on fossil fuels by providing biofuels, plastics and industrial chemicals. Gaining an understanding of the relevant functions of the wide range of microbes that produce, consume and modify hydrocarbons and related compounds will be key to responding to these challenges. This comprehensive collection of current and emerging protocols will facilitate acquisition of this understanding and exploitation of useful activities of such microbes.

This Volume presents generic protocols for wet experimental and computer-based systems and synthetic biology approaches relevant to the field of hydrocarbon and lipid microbiology.

This Volume provides protocols for the biochemical analysis of hydrocarbon- and lipid-relevant products, cell components and activities of microbes that interact with hydrophobic compounds.

This Volume covers protocols for in-silico approaches to hydrocarbon microbiology, including the selection and use of appropriate statistical tools for experimental design replication, data analysis, and computer-assisted approaches to data storage, management and utilisation. The application of algorithms to analyse the composition and function of microbial communities is presented, as are prediction tools for biodegradation and protein interactions. The basics of a major open-source programming language, Python, are explained. Protocols for calculating reaction kinetics and thermodynamics are presented, and modelling the environmental fate of hydrocarbons during bioremediation is explained. With the exception of molecular biology studies of molecular interactions, the use of statistics is absolutely essential for both experimental design and data analysis in microbiological research, and indeed in the biomedical sciences in general. Moreover, studies of highly varying systems call for the modelling and/or application of theoretical frameworks. Thus, while two protocols in this Volume are specific to hydrocarbon microbiology, the others are generic, and as such will be of use to researchers investigating a broad range of topics in microbiology and the biomedical sciences in general.Hydrocarbon and Lipid Microbiology ProtocolsThere are tens of thousands of structurally different hydrocarbons, hydrocarbon derivatives and lipids, and a wide array of these molecules are required for cells to function. The global hydrocarbon cycle, which is largely driven by microorganisms, has a major impact on our environment and climate. Microbes are responsible for cleaning up the environmental pollution caused by the exploitation of hydrocarbon reservoirs and will also be pivotal in reducing our reliance on fossil fuels by providing biofuels, plastics and industrial chemicals. Gaining an understanding of the relevant functions of the wide range of microbes that produce, consume and modify hydrocarbons and related compounds will be key to responding to these challenges. This comprehensive collection of current and emerging protocols will facilitate acquisition of this understanding and exploitation of useful activities of such microbes.