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Full Description
Metabolic Flux Analysis: Methods and Protocols opens up the field of metabolic flux analysis to those who want to start a new flux analysis project but are overwhelmed by the complexity of the approach. Metabolic flux analysis emerged from the current limitation for the prediction of metabolic fluxes from a measured inventory of the cell. Divided into convenient thematic parts, topics in this essential volume include the fundamental characteristics of the underlying networks, the application of quantitative metabolite data and thermodynamic principles to constrain the solution space for flux balance analysis (FBA), the experimental toolbox to conduct different types of flux analysis experiments, the processing of data from 13C experiments and three chapters that summarize some recent key findings. Written in the 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 protocols and notes on troubleshooting and avoiding known pitfalls.
Authoritative and easily accessible, Metabolic Flux Analysis: Methods and Protocols presents protocols that cover a range of relevant organisms currently used in the field, providing a solid basis to anybody interested in the field of metabolic flux analysis.
Contents
Stoichiometric Modelling of Microbial Metabolism.- Tapping the Wealth of Microbial Data in High-Throughput Metabolic Model Reconstruction.- Constraining the Flux Space using Thermodynamics and Integration of Metabolomics Data.- NExT - Integration of Thermodynamic Constraints and Metabolomics Data into a Metabolic Network.- Customization of 13C-MFA Strategy According to Cell Culture System.- Quantitative Metabolomics Using ID-MS.- Determining the Biomass Composition of a Sponge Holobiont for Flux Analysis.- Successful Down-Sizing for High-Throughput 13C-MFA Applications. Labelling Analysis for 13C MFA Using NMR Spectroscopy.- GC-MS Based 13C Metabolic Flux Analysis.- Dynamic Cell Respiration & CO2 Labeling Analysis Using Membrane-Inlet Mass Spectrometry.- Correction of MS Data for Naturally Occurring Isotopes in Isotope Labelling Experiments.- Steady-State 13C Fluxomics Using OpenFLUX.- Flux Visualization Using VANTED/FluxMap.- Metabolic Flux Analysis for Escherichia coli by Flux Balance Analysis.- 13C-Metabolic Flux Analysis for Escherichia coli.- 3C-Based Metabolic Flux Analysis of Recombinant Pichia pastoris.
