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This volume provides up-to-date and novel techniques for various screening technologies currently used in metagenomics and related areas. Starting with DNA/RNA isolation from environmental samples, the book continues by delving into areas such as current methods used to isolate DNA and construct metagenomic libraries, establishment of metagenome libraries in non-E. coli hosts, and topics like function-driven mining of metagenomic DNA, screening and analyzing protocols for a wide array of different genes encoding enzymes, bacterial viruses and much more. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Metagenomics: Methods and Protocols, Third Edition provides a comprehensive collection of up-to-date metagenome protocols and tools for the recovery of many major types of biocatalysts and allows for the easy setup of these screens in microbiology laboratories.
Sommario
Construction of Small-Insert and Large-Insert Metagenomic Libraries.- Extraction of Total DNA and RNA from Marine Filter Samples and Generation of a Universal cDNA as Universal Template for Marker Gene Studies.- Functional Metagenomics as a Tool to Tap into Natural Diversity of Valuable Biotechnological Compounds.- Functional and Sequence-Specific Screening Protocols for the Detection of Novel Antimicrobial Resistance Genes in Metagenomic DNA.- Identification of PKS Gene Clusters from Metagenomic Libraries Using a Next Generation Sequencing Approach.- Activity-Based Screening of Metagenomic Fosmid Libraries for Hydrogen-Uptake Enzymes.- Functional Metagenomics Approach for the Discovery of Novel Genes Encoding Phosphatase Activity.- Isolation of Genes Encoding Carbon Metabolism Pathways from Complex Microbial Communities.- Screening Metagenomes for Algae Cell Wall Carbohydrates Degrading Hydrolases in Enrichment Cultures.- The PET-Degrading Potential of Global Metagenomes: From InSilico Mining to Active Enzymes.- High-Throughput Screening for Thermostable Polyester Hydrolases.- Metagenomic Screening of a Novel PET Esterase via In Vitro Expression System.- Assigning Functions of Unknown Enzymes by High Throughput Enzyme Characterization.- Isolation of a Host-Confined Phage Metagenome Allows the Detection of Phages Both Capable and Incapable of Plaque Formation.- CRISPR-Cas9 Shaped Viral Metagenomes Associated with Bacillus subtilis.- Cloning and Expression of Metagenomic DNA in Streptomyces lividans and Its Subsequent Fermentation for Optimized Production.- DNA-, RNA-, and Protein-Based Stable-Isotope Probing for High-Throughput Biomarker Analysis of Active Microorganisms.
