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...

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 Protocols

There are tens of...

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. It presents detailed protocols for the functional screening of enzymes acting on greasy molecules (i.e. lipases, esterases), including assays for enantioselective biocatalysts, as well as approaches for protein display technologies. Protocols for improving fuel quality and production of biofuel and lipids in different hosts (bacteria, algae, yeast) are also provided. The...

This Volume presents applications of hydrocarbon microbiology in the context of environmental pollutant degradation, covering pollutants such as petroleum and related wastes (i.e. oil sludge), biofuels, lipid-rich wastes, chlorinated solvents and BTEX, in several environments (marine, soil, groundwater). The approaches presented range from laboratory experiments and treatment in reactors to field applications. Two chapters highlight innovative approaches to address relevant questions in pollutant degradation, such as low environmental concentrations of pollutants, and the biodegradation of...

This Volume describes methods for simulating natural environments by using reproducible and controllable meso- and microcosm experiments to analyse hydrocarbon-degrading microorganisms and to test hypotheses. It presents important aspects of the preparation of experimental set-ups, hydrocarbon application and sampling, and features protocols for experiments with different types of samples, such as biofilms, aquatic systems (e.g. groundwater, streams), and sediments, including oscillating oxic-anoxic mesocosms, as well as for in-situ experimentation in subtidal and deep sediments. Two...

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. Methods using different probe and cell types, tissues and organisms, such as mammalians, fish, amphibians (including lampbrush-chromosomes), insects, plants and microorganisms are described in 57 chapters. In addition to multicolor FISH procedures and special applications such as the characterization of marker chromosomes, breakpoints, cryptic aberrations, nuclear...