{"product_id":"molecular-tools-for-the-detection-and-quantification-of-toxigenic-cyanobacteria-isbn-9781119332107","title":"Molecular Tools for the Detection and Quantification of Toxigenic Cyanobacteria","description":"\u003cp\u003e\u003cb\u003eA guide to state-of-the-art molecular tools for \u003c\/b\u003e\u003cb\u003emonitoring and managing the toxigenicity of cyanobacteria\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003eRunaway eutrophication and climate change has made the monitoring and management of toxigenic organisms in the world’s bodies of water more urgent than ever. In order to influence public policy regarding the detection and quantification of those organisms, it is incumbent upon scientists to raise the awareness of policy makers concerning the increased occurrence of toxigenic cyanobacteria and the threats they pose. As molecular methods can handle many samples in short time and help identify toxigenic organisms, they are reliable, cost-effective tools available for tracking toxigenic cyanobacteria worldwide. This volume arms scientists with the tools they need to track toxigenicity in surface waters and food supplies and, hopefully, to develop new techniques for managing the spread of toxic cyanobacteria.\u003c\/p\u003e \u003cp\u003eThis handbook offers the first comprehensive treatment of molecular tools for monitoring toxigenic cyanobacteria. Growing out of the findings of the landmark European Cooperation in Science and Technology Cyanobacteria project (CYANOCOST), it provides detailed, practical coverage of the full array of available molecular tools and protocols, from water sampling, nucleic acid extraction, and downstream analysis—including PCR and qPCR based methods—to  genotyping (DGGE), diagnostic microarrays, and community characterization using next-gen sequencing techniques.\u003c\/p\u003e \u003cul\u003e \u003cli\u003eOffers an overview of the latest trends in the field, while providing a foundation for understanding and applying the tools and techniques described\u003c\/li\u003e \u003cli\u003eProvides detailed coverage of the full range of molecular tools currently available, with expert guidance on the analysis and interpretation of results\u003c\/li\u003e \u003cli\u003eIncludes step-by-step guidance on standard operational procedures, including molecular tests used in environmental monitoring, with individual chapters devoted to each procedure\u003c\/li\u003e \u003cli\u003eComplements the published \u003ci\u003eHandbook of Cyanobacterial Monitoring and Cyanotoxin Analysis\u003c\/i\u003e from the CyanoCOST project\u003c\/li\u003e \u003c\/ul\u003e \u003cp\u003eThis handbook is an indispensable working resource for scientists, lab technicians, and water management professionals and an excellent text\/reference for graduate students and supervisors who use molecular tools. It will also be of great value to environmental health and protection officials and policy makers.\u003c\/p\u003e \u003cp\u003eList of Contributors xix\u003c\/p\u003e \u003cp\u003eAbout the Editors xxiii\u003c\/p\u003e \u003cp\u003eAbout the Book xxvii\u003c\/p\u003e \u003cp\u003ePreface xxix\u003c\/p\u003e \u003cp\u003eAcknowledgments xxxi\u003c\/p\u003e \u003cp\u003e1 Introduction 1\u003cbr\u003e\u003ci\u003eRainer Kurmayer, Kaarina Sivonen, and Nico Salmaso\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e1.1 A Brief Historical Overview 1\u003c\/p\u003e \u003cp\u003e1.2 The Genetic Basis of Toxin Production 2\u003c\/p\u003e \u003cp\u003e1.2.1 Microcystin and Nodularin 2\u003c\/p\u003e \u003cp\u003e1.2.2 Cylindrospermopsin 5\u003c\/p\u003e \u003cp\u003e1.2.3 Saxitoxin 6\u003c\/p\u003e \u003cp\u003e1.2.4 Anatoxin 8\u003c\/p\u003e \u003cp\u003e1.3 Application of Molecular Tools 8\u003c\/p\u003e \u003cp\u003e1.4 Laboratory Safety Issues 13\u003c\/p\u003e \u003cp\u003e1.5 References 14\u003c\/p\u003e \u003cp\u003e2 Sampling and Metadata 19\u003cbr\u003e\u003ci\u003eRainer Kurmayer, Guntram Christiansen, Konstantinos Kormas, Wim Vyverman, Elie Verleyen, Vitor Ramos, Vitor Vasconcelos, and Nico Salmaso\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e2.1 Introduction 19\u003c\/p\u003e \u003cp\u003e2.2 Handling of Samples 20\u003c\/p\u003e \u003cp\u003e2.3 Sample Contamination 21\u003c\/p\u003e \u003cp\u003e2.4 Sampling 21\u003c\/p\u003e \u003cp\u003e2.4.1 Quantitative Depth-Integrated and Discrete Sampling 21\u003c\/p\u003e \u003cp\u003e2.4.2 Qualitative Plankton Net Sampling 22\u003c\/p\u003e \u003cp\u003e2.4.3 Surface (Scum Material) Sampling 22\u003c\/p\u003e \u003cp\u003e2.4.4 Benthic (Terrestrial) Cyanobacteria Sampling 22\u003c\/p\u003e \u003cp\u003e2.4.5 Food Supplement Sampling 22\u003c\/p\u003e \u003cp\u003e2.4.6 Isolation of Single Colonies\/Filaments 22\u003c\/p\u003e \u003cp\u003e2.5 Subsampling Food Supplement Samples 23\u003c\/p\u003e \u003cp\u003e2.6 Sampling of Nucleic Acids 23\u003c\/p\u003e \u003cp\u003e2.7 General Conclusions 24\u003c\/p\u003e \u003cp\u003e2.8 References 24\u003c\/p\u003e \u003cp\u003eSOP 2.1 Sampling and Filtration (DNA) 26\u003cbr\u003e\u003ci\u003eRainer Kurmayer and Konstantinos Kormas\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 2.1.1 Introduction 26\u003c\/p\u003e \u003cp\u003eSOP 2.1.2 Experimental 26\u003c\/p\u003e \u003cp\u003eSOP 2.1.3 Procedure 27\u003c\/p\u003e \u003cp\u003eSOP 2.1.4 Notes 28\u003c\/p\u003e \u003cp\u003eSOP 2.1.5 References 29\u003c\/p\u003e \u003cp\u003eSOP 2.2 Sampling of Benthic Cyanobacteria 29\u003cbr\u003e\u003ci\u003eWim Vyverman and Elie Verleyen\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 2.2.1 Introduction 29\u003c\/p\u003e \u003cp\u003eSOP 2.2.2 Experimental 30\u003c\/p\u003e \u003cp\u003eSOP 2.2.3 Procedure 30\u003c\/p\u003e \u003cp\u003eSOP 2.2.4 Notes 31\u003c\/p\u003e \u003cp\u003eSOP 2.2.5 References 31\u003c\/p\u003e \u003cp\u003eSOP 2.3 Isolation of Single Cyanobacteria Colonies\/Filaments 32\u003c\/p\u003e \u003cp\u003eRainer Kurmayer\u003c\/p\u003e \u003cp\u003eSOP 2.3.1 Introduction 32\u003c\/p\u003e \u003cp\u003eSOP 2.3.2 Experimental 32\u003c\/p\u003e \u003cp\u003eSOP 2.3.3 Procedure 33\u003c\/p\u003e \u003cp\u003eSOP 2.3.4 Notes 33\u003c\/p\u003e \u003cp\u003eSOP 2.3.5 References 33\u003c\/p\u003e \u003cp\u003eSOP 2.4 Sampling Food Supplements 34\u003cbr\u003e\u003ci\u003eVitor Ramos, Cristiana Moreira, and Vitor Vasconcelos\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 2.4.1 Introduction 34\u003c\/p\u003e \u003cp\u003eSOP 2.4.2 Experimental 35\u003c\/p\u003e \u003cp\u003eSOP 2.4.3 Procedure (Fig. 8.3) 35\u003c\/p\u003e \u003cp\u003eSOP 2.4.4 Notes 36\u003c\/p\u003e \u003cp\u003eSOP 2.4.5 References 36\u003c\/p\u003e \u003cp\u003eSOP 2.5 Sampling and Filtration (RNA) 37\u003cbr\u003e\u003ci\u003eRainer Kurmayer and Guntram Christiansen\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 2.5.1 Introduction 37\u003c\/p\u003e \u003cp\u003eSOP 2.5.2 Experimental 37\u003c\/p\u003e \u003cp\u003eSOP 2.5.3 Procedure 38\u003c\/p\u003e \u003cp\u003eSOP 2.5.4 Notes 38\u003c\/p\u003e \u003cp\u003eSOP 2.5.5 References 38\u003c\/p\u003e \u003cp\u003eSOP 2.6 Sampling of Abiotic and Biotic Data and Recording Metadata 39\u003cbr\u003e\u003ci\u003eElie Verleyen, Maxime Sweetlove, Dagmar Obbels, and Wim Vyverman\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 2.6.1 Introduction 39\u003c\/p\u003e \u003cp\u003eSOP 2.6.2 Experimental 39\u003c\/p\u003e \u003cp\u003eSOP 2.6.3 Type of Metadata and Additional Biotic and Abiotic Data 40\u003c\/p\u003e \u003cp\u003eSOP 2.6.4 Notes 41\u003c\/p\u003e \u003cp\u003eSOP 2.6.5 References 42\u003c\/p\u003e \u003cp\u003e3 Isolation, Purification, and Cultivation of Toxigenic Cyanobacteria 43\u003cbr\u003e\u003ci\u003eSigrid Haande, Iwona Jasser, Muriel Gugger, Camilla H.C. Hagman, Annick Wilmotte, and Andreas Ballot\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e3.1 Introduction 43\u003c\/p\u003e \u003cp\u003e3.2 Methodical Principles for Cyanobacterial Isolation, Purification, and Cultivation 44\u003c\/p\u003e \u003cp\u003e3.2.1 Sampling, Identification, and Treatments Prior to the Isolation of Cyanobacteria 44\u003c\/p\u003e \u003cp\u003e3.2.2 Traditional Techniques for the Isolation and Purification of Cyanobacteria 45\u003c\/p\u003e \u003cp\u003e3.2.3 Culture Media Preparation 47\u003c\/p\u003e \u003cp\u003e3.2.4 Cultivation Conditions 48\u003c\/p\u003e \u003cp\u003e3.3 General Conclusions 49\u003c\/p\u003e \u003cp\u003e3.4 References 49\u003c\/p\u003e \u003cp\u003eSOP 3.1 Isolation, Purification, and Clonal Isolate Testing 51\u003cbr\u003e\u003ci\u003eSigrid Haande, Camilla H.C. Hagman, and Andreas Ballot\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 3.1.1 Introduction 51\u003c\/p\u003e \u003cp\u003eSOP 3.1.2 Experimental 51\u003c\/p\u003e \u003cp\u003eSOP 3.1.3 Procedure 52\u003c\/p\u003e \u003cp\u003eSOP 3.1.4 Notes 54\u003c\/p\u003e \u003cp\u003eSOP 3.1.5 References 54\u003c\/p\u003e \u003cp\u003eSOP 3.2 Isolation of Picocyanobacterial Cells by Flow Cytometer (FCM) Sorting 55\u003cbr\u003e\u003ci\u003eEwa Koz³owska and Iwona Jasser\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 3.2.1 Introduction 55\u003c\/p\u003e \u003cp\u003eSOP 3.2.2 Experimental 56\u003c\/p\u003e \u003cp\u003eSOP 3.2.3 Procedure 56\u003c\/p\u003e \u003cp\u003eSOP 3.2.4 Notes 58\u003c\/p\u003e \u003cp\u003eSOP 3.2.5 References 59\u003c\/p\u003e \u003cp\u003eSOP 3.3 Axenization 60\u003cbr\u003e\u003ci\u003eMuriel Gugger\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 3.3.1 Introduction 60\u003c\/p\u003e \u003cp\u003eSOP 3.3.2 Experimental 60\u003c\/p\u003e \u003cp\u003eSOP 3.3.3 Procedure 61\u003c\/p\u003e \u003cp\u003eSOP 3.3.4 Notes 63\u003c\/p\u003e \u003cp\u003eSOP 3.3.5 References 63\u003c\/p\u003e \u003cp\u003eSOP 3.4 Culture Media (Solid and Liquid) 64\u003cbr\u003e\u003ci\u003eSigrid Haande, Camilla H.C. Hagman, and Andreas Ballot\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 3.4.1 Introduction 64\u003c\/p\u003e \u003cp\u003eSOP 3.4.2 Experimental 64\u003c\/p\u003e \u003cp\u003eSOP 3.4.3 Procedure 65\u003c\/p\u003e \u003cp\u003eSOP 3.4.4 Notes 68\u003c\/p\u003e \u003cp\u003eSOP 3.4.5 References 68\u003c\/p\u003e \u003cp\u003eSOP 3.5 Strain Maintenance (Living Cultures) 69\u003cbr\u003e\u003ci\u003eSigrid Haande, Camilla H.C. Hagman, and Andreas Ballot\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 3.5.1 Introduction 69\u003c\/p\u003e \u003cp\u003eSOP 3.5.2 Experimental 69\u003c\/p\u003e \u003cp\u003eSOP 3.5.3 Procedure 70\u003c\/p\u003e \u003cp\u003eSOP 3.5.4 Notes 72\u003c\/p\u003e \u003cp\u003eSOP 3.5.5 References 73\u003c\/p\u003e \u003cp\u003eSOP 3.6 Cryopreservation and Recovery 73\u003cbr\u003e\u003ci\u003eMuriel Gugger\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 3.6.1 Introduction 73\u003c\/p\u003e \u003cp\u003eSOP 3.6.2 Experimental 74\u003c\/p\u003e \u003cp\u003eSOP 3.6.3 Procedure 75\u003c\/p\u003e \u003cp\u003eSOP 3.6.4 Notes 78\u003c\/p\u003e \u003cp\u003eSOP 3.6.5 References 78\u003c\/p\u003e \u003cp\u003e4 Taxonomic Identification of Cyanobacteria by a Polyphasic Approach 79\u003cbr\u003e\u003ci\u003eAnnick Wilmotte, H. Dail Laughinghouse IV, Camilla Capelli, Rosmarie Rippka, and Nico Salmaso\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e4.1 Introduction 79\u003c\/p\u003e \u003cp\u003e4.2 Nomenclature and Classification of Cyanobacteria 82\u003c\/p\u003e \u003cp\u003e4.3 Microscopy 84\u003c\/p\u003e \u003cp\u003e4.3.1 Light Microscopy 84\u003c\/p\u003e \u003cp\u003e4.3.2 Autofluorescence Microscopy 86\u003c\/p\u003e \u003cp\u003e4.4 Molecular Markers: Single Loci 87\u003c\/p\u003e \u003cp\u003e4.5 Molecular Markers: Multiple Loci 94\u003c\/p\u003e \u003cp\u003e4.5.1 Multilocus Sequence Typing (MLST) and Multilocus Sequence Analysis (MLSA) 94\u003c\/p\u003e \u003cp\u003e4.5.2 Genome-Based Extension of MLST and MLSA 96\u003c\/p\u003e \u003cp\u003e4.6 Molecular Typing Methods Based on Gel Electrophoresis 96\u003c\/p\u003e \u003cp\u003e4.7 Denaturing Gradient Gel Electrophoresis (DGGE) 97\u003c\/p\u003e \u003cp\u003e4.8 Taxonomic and Molecular Databases 97\u003c\/p\u003e \u003cp\u003e4.9 The Polyphasic Approach 98\u003c\/p\u003e \u003cp\u003e4.10 Final Considerations 105\u003c\/p\u003e \u003cp\u003e4.11 References 106\u003c\/p\u003e \u003cp\u003eSOP 4.1 Taxonomic Identification by Light Microscopy 120\u003cbr\u003e\u003ci\u003eNico Salmaso, Rosmarie Rippka, and Annick Wilmotte\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 4.1.1 Introduction 120\u003c\/p\u003e \u003cp\u003eSOP 4.1.2 Experimental 121\u003c\/p\u003e \u003cp\u003eSOP 4.1.3 References 124\u003c\/p\u003e \u003cp\u003eSOP 4.2 Polyphasic Approach on Cyanobacterial Strains 125\u003cbr\u003e\u003ci\u003eNico Salmaso, Camilla Capelli, Rosmarie Rippka, and Annick Wilmotte\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 4.2.1 Introduction 125\u003c\/p\u003e \u003cp\u003eSOP 4.2.2 Experimental 126\u003c\/p\u003e \u003cp\u003eSOP 4.2.3 References 131\u003c\/p\u003e \u003cp\u003e5 Nucleic Acid Extraction 135\u003cbr\u003e\u003ci\u003eElke Dittmann, Anne Rantala-Ylinen, Vitor Ramos, Vitor Vasconcelos, Guntram Christiansen, and Rainer Kurmayer\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e5.1 Introduction 135\u003c\/p\u003e \u003cp\u003e5.2 Specific Extraction Procedures and Storage 137\u003c\/p\u003e \u003cp\u003e5.2.1 DNA Extraction from Laboratory Strains 137\u003c\/p\u003e \u003cp\u003e5.2.2 DNA Extraction from Field Samples 137\u003c\/p\u003e \u003cp\u003e5.2.3 DNA Extraction from Food Supplements 137\u003c\/p\u003e \u003cp\u003e5.2.4 RNA Extraction from Laboratory Strains 138\u003c\/p\u003e \u003cp\u003e5.2.5 RNA Extraction from Field Samples 138\u003c\/p\u003e \u003cp\u003e5.2.6 Single Colony and Filament Analysis 138\u003c\/p\u003e \u003cp\u003e5.2.7 Whole Genome Amplification 139\u003c\/p\u003e \u003cp\u003e5.2.8 Nucleic Acid Storage 139\u003c\/p\u003e \u003cp\u003e5.3 References 139\u003c\/p\u003e \u003cp\u003eSOP 5.1 Standard DNA Isolation Technique for Cyanobacteria 140\u003cbr\u003e\u003ci\u003eElke Dittmann\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 5.1.1 Introduction 140\u003c\/p\u003e \u003cp\u003eSOP 5.1.2 Experimental 140\u003c\/p\u003e \u003cp\u003eSOP 5.1.3 Procedure 141\u003c\/p\u003e \u003cp\u003eSOP 5.1.4 Notes 141\u003c\/p\u003e \u003cp\u003eSOP 5.1.5 References 142\u003c\/p\u003e \u003cp\u003eSOP 5.2 DNA Isolation Protocol for Cyanobacteria with Extensive Mucilage 143\u003cbr\u003e\u003ci\u003eGuntram Christiansen, Elisabeth Entfellner, and Rainer Kurmayer\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 5.2.1 Introduction 143\u003c\/p\u003e \u003cp\u003eSOP 5.2.2 Experimental 143\u003c\/p\u003e \u003cp\u003eSOP 5.2.3 Procedure 144\u003c\/p\u003e \u003cp\u003eSOP 5.2.4 Notes 145\u003c\/p\u003e \u003cp\u003eSOP 5.2.5 References 145\u003c\/p\u003e \u003cp\u003eSOP 5.3 Quantitative DNA Isolation from Filters 145\u003cbr\u003e\u003ci\u003eRainer Kurmayer\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 5.3.1 Introduction 146\u003c\/p\u003e \u003cp\u003eSOP 5.3.2 Experimental 146\u003c\/p\u003e \u003cp\u003eSOP 5.3.3 Procedure 147\u003c\/p\u003e \u003cp\u003eSOP 5.3.4 Notes 148\u003c\/p\u003e \u003cp\u003eSOP 5.3.5 References 148\u003c\/p\u003e \u003cp\u003eSOP 5.4 Genomic DNA Extraction from Single Filaments\/Colonies for Multiple PCR Analyses 149\u003cbr\u003e\u003ci\u003eGuntram Christiansen, Chen Qin, and Rainer Kurmayer\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 5.4.1 Introduction 149\u003c\/p\u003e \u003cp\u003eSOP 5.4.2 Experimental 149\u003c\/p\u003e \u003cp\u003eSOP 5.4.3 Procedure 150\u003c\/p\u003e \u003cp\u003eSOP 5.4.4 Notes 151\u003c\/p\u003e \u003cp\u003eSOP 5.4.5 References 151\u003c\/p\u003e \u003cp\u003eSOP 5.5 Whole Genome Amplification Using Bacteriophage Phi29 DNA Polymerase 151\u003cbr\u003e\u003ci\u003eGuntram Christiansen and Rainer Kurmayer\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 5.5.1 Introduction 151\u003c\/p\u003e \u003cp\u003eSOP 5.5.2 Experimental 152\u003c\/p\u003e \u003cp\u003eSOP 5.5.3 Procedure 152\u003c\/p\u003e \u003cp\u003eSOP 5.5.4 Notes 152\u003c\/p\u003e \u003cp\u003eSOP 5.5.5 Reference 153\u003c\/p\u003e \u003cp\u003eSOP 5.6 DNA Extraction from Food Supplements 153\u003cbr\u003e\u003ci\u003eVitor Ramos, Cristiana Moreira, and Vitor Vasconcelos\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 5.6.1 Introduction 153\u003c\/p\u003e \u003cp\u003eSOP 5.6.2 Experimental 153\u003c\/p\u003e \u003cp\u003eSOP 5.6.3 Procedure 154\u003c\/p\u003e \u003cp\u003eSOP 5.6.4 Notes 155\u003c\/p\u003e \u003cp\u003eSOP 5.6.5 References 156\u003c\/p\u003e \u003cp\u003eSOP 5.7 RNA Extraction from Cyanobacteria 156\u003cbr\u003e\u003ci\u003eGuntram Christiansen and Rainer Kurmayer\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 5.7.1 Introduction 156\u003c\/p\u003e \u003cp\u003eSOP 5.7.2 Experimental 156\u003c\/p\u003e \u003cp\u003eSOP 5.7.3 Procedure 158\u003c\/p\u003e \u003cp\u003eSOP 5.7.4 Notes 158\u003c\/p\u003e \u003cp\u003eSOP 5.7.5 References 159\u003c\/p\u003e \u003cp\u003eSOP 5.8 cDNA Synthesis 159\u003cbr\u003e\u003ci\u003eGuntram Christiansen and Rainer Kurmayer\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 5.8.1 Introduction 159\u003c\/p\u003e \u003cp\u003eSOP 5.8.2 Experimental 159\u003c\/p\u003e \u003cp\u003eSOP 5.8.3 Procedure 160\u003c\/p\u003e \u003cp\u003eSOP 5.8.4 Notes 161\u003c\/p\u003e \u003cp\u003eSOP 5.8.5 References 161\u003c\/p\u003e \u003cp\u003e6 Conventional PCR 163\u003cbr\u003e\u003ci\u003eElke Dittmann, Anne Rantala-Ylinen, Kaarina Sivonen, Ilona Ga²ga³a, Joanna\u003c\/i\u003e \u003ci\u003eMankiewicz-Boczek, Samuel Cirés, Andreas Ballot, Guntram Christiansen,\u003c\/i\u003e \u003ci\u003eRainer Kurmayer, Vitor Ramos, Vitor Vasconcelos, and Martin Saker\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e6.1 Introduction 163\u003c\/p\u003e \u003cp\u003e6.2 Principle of PCR and Available Enzymes 164\u003c\/p\u003e \u003cp\u003e6.2.1 Primer Development 165\u003c\/p\u003e \u003cp\u003e6.2.2 Setup of PCR Conditions for DNA and Single Colony Analysis 168\u003c\/p\u003e \u003cp\u003e6.2.3 Gel Electrophoresis and Documentation 168\u003c\/p\u003e \u003cp\u003e6.2.4 Troubleshooting of PCR Results 168\u003c\/p\u003e \u003cp\u003e6.2.5 PCR Product Downstream Processing (RFLP, Cloning, Sequencing) 169\u003c\/p\u003e \u003cp\u003e6.3 Special Notes 170\u003c\/p\u003e \u003cp\u003e6.4 References 170\u003c\/p\u003e \u003cp\u003eSOP 6.1 PCR Detection of Microcystin Biosynthesis Genes Combined with RFLP Differentiation of the Producing Genus 172\u003cbr\u003e\u003ci\u003eElke Dittmann\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 6.1.1 Introduction 172\u003c\/p\u003e \u003cp\u003eSOP 6.1.2 Experimental 172\u003c\/p\u003e \u003cp\u003eSOP 6.1.3 Procedure 173\u003c\/p\u003e \u003cp\u003eSOP 6.1.4 Notes 174\u003c\/p\u003e \u003cp\u003eSOP 6.1.5 Reference 174\u003c\/p\u003e \u003cp\u003eSOP 6.2 PCR Detection of Microcystin and Nodularin Biosynthesis Genes in the Cyanobacterial Orders Oscillatoriales, Chroococcales, Stigonematales, and Nostocales 175\u003cbr\u003e\u003ci\u003eElke Dittmann, Joanna Mankiewicz-Boczek, and Ilona Ga²ga³a\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 6.2.1 Introduction 175\u003c\/p\u003e \u003cp\u003eSOP 6.2.2 Experimental 175\u003c\/p\u003e \u003cp\u003eSOP 6.2.3 Procedure 177\u003c\/p\u003e \u003cp\u003eSOP 6.2.4 Notes 177\u003c\/p\u003e \u003cp\u003eSOP 6.2.5 References 178\u003c\/p\u003e \u003cp\u003eSOP 6.3 Genus-Specific PCR Detection of Microcystin Biosynthesis Genes in Anabaena\/Nodularia and Microcystis and Planktothrix, Respectively 179\u003cbr\u003e\u003ci\u003eAnne Rantala-Ylinen and Kaarina Sivonen\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 6.3.1 Introduction 179\u003c\/p\u003e \u003cp\u003eSOP 6.3.2 Experimental 179\u003c\/p\u003e \u003cp\u003eSOP 6.3.3 Procedure 181\u003c\/p\u003e \u003cp\u003eSOP 6.3.4 Notes 181\u003c\/p\u003e \u003cp\u003eSOP 6.3.5 References 181\u003c\/p\u003e \u003cp\u003eSOP 6.4 PCR Detection of Anatoxin Biosynthesis Genes Combined with RFLP Differentiation of the Producing Genus 182\u003cbr\u003e\u003ci\u003eAnne Rantala-Ylinen and Kaarina Sivonen\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 6.4.1 Introduction 182\u003c\/p\u003e \u003cp\u003eSOP 6.4.2 Experimental 182\u003c\/p\u003e \u003cp\u003eSOP 6.4.3 Procedure 183\u003c\/p\u003e \u003cp\u003eSOP 6.4.4 Notes 184\u003c\/p\u003e \u003cp\u003eSOP 6.4.5 Reference 184\u003c\/p\u003e \u003cp\u003eSOP 6.5 PCR Detection of the Saxitoxin Biosynthesis Genes, sxtA, sxtX, sxtH, sxtG, and sxtI 185\u003cbr\u003e\u003ci\u003eAndreas Ballot and Samuel Cirés\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 6.5.1 Introduction 185\u003c\/p\u003e \u003cp\u003eSOP 6.5.2 Experimental 187\u003c\/p\u003e \u003cp\u003eSOP 6.5.3 Procedure 187\u003c\/p\u003e \u003cp\u003eSOP 6.5.4 Notes 188\u003c\/p\u003e \u003cp\u003eSOP 6.5.5 References 189\u003c\/p\u003e \u003cp\u003eSOP 6.6 PCR Detection of the Cylindrospermopsin Biosynthesis Gene cyrJ 189\u003cbr\u003e\u003ci\u003eSamuel Cirés and Andreas Ballot\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 6.6.1 Introduction 189\u003c\/p\u003e \u003cp\u003eSOP 6.6.2 Experimental 190\u003c\/p\u003e \u003cp\u003eSOP 6.6.3 Procedure 191\u003c\/p\u003e \u003cp\u003eSOP 6.6.4 Notes 191\u003c\/p\u003e \u003cp\u003eSOP 6.6.5 References 192\u003c\/p\u003e \u003cp\u003eSOP 6.7 PCR from Single Filament of Toxigenic Planktothrix 193\u003cbr\u003e\u003ci\u003eQin Chen, Guntram Christiansen, and Rainer Kurmayer\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 6.7.1 Introduction 193\u003c\/p\u003e \u003cp\u003eSOP 6.7.2 Experimental 193\u003c\/p\u003e \u003cp\u003eSOP 6.7.3 Procedure 194\u003c\/p\u003e \u003cp\u003eSOP 6.7.4 Notes 195\u003c\/p\u003e \u003cp\u003eSOP 6.7.5 References 195\u003c\/p\u003e \u003cp\u003eSOP 6.8 Analysis of Microcystin Biosynthesis Gene Subpopulation Variability in Planktothrix 196\u003cbr\u003e\u003ci\u003eRainer Kurmayer\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 6.8.1 Introduction 196\u003c\/p\u003e \u003cp\u003eSOP 6.8.2 Experimental 196\u003c\/p\u003e \u003cp\u003eSOP 6.8.3 Procedure 197\u003c\/p\u003e \u003cp\u003eSOP 6.8.4 Notes 197\u003c\/p\u003e \u003cp\u003eSOP 6.8.5 References 198\u003c\/p\u003e \u003cp\u003eSOP 6.9 PCR Detection of Microcystin Biosynthesis Genes from Food\u003c\/p\u003e \u003cp\u003eSupplements 199\u003cbr\u003e\u003ci\u003eVitor Ramos, Cristiana Moreira, and Vitor Vasconcelos\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 6.9.1 Introduction 199\u003c\/p\u003e \u003cp\u003eSOP 6.9.2 Experimental 199\u003c\/p\u003e \u003cp\u003eSOP 6.9.3 Procedure 201\u003c\/p\u003e \u003cp\u003eSOP 6.9.4 Notes 202\u003c\/p\u003e \u003cp\u003eSOP 6.9.5 References 203\u003c\/p\u003e \u003cp\u003e7 Quantitative PCR 205\u003cbr\u003e\u003ci\u003eAnne Rantala-Ylinen, Henna Savela, Kaarina Sivonen, and Rainer Kurmayer\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e7.1 Introduction 205\u003c\/p\u003e \u003cp\u003e7.2 Primer\/Probe Design 206\u003c\/p\u003e \u003cp\u003e7.3 Optimization 208\u003c\/p\u003e \u003cp\u003e7.4 Absolute Quantification 208\u003c\/p\u003e \u003cp\u003e7.5 Relative Quantification 209\u003c\/p\u003e \u003cp\u003e7.6 Calibration of qPCR Results 209\u003c\/p\u003e \u003cp\u003e7.7 General Conclusions 210\u003c\/p\u003e \u003cp\u003e7.8 References 210\u003c\/p\u003e \u003cp\u003eSOP 7.1 Optimization of qPCR Assays 211\u003cbr\u003e\u003ci\u003eRainer Kurmayer\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 7.1.1 Introduction 211\u003c\/p\u003e \u003cp\u003eSOP 7.1.2 Experimental 211\u003c\/p\u003e \u003cp\u003eSOP 7.1.3 Procedure 212\u003c\/p\u003e \u003cp\u003eSOP 7.1.4 Notes 213\u003c\/p\u003e \u003cp\u003eSOP 7.1.5 References 213\u003c\/p\u003e \u003cp\u003eSOP 7.2 Calibration of qPCR Results 214\u003cbr\u003e\u003ci\u003eRainer Kurmayer\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 7.2.1 Introduction 214\u003c\/p\u003e \u003cp\u003eSOP 7.2.2 Experimental 214\u003c\/p\u003e \u003cp\u003eSOP 7.2.3 Procedure 215\u003c\/p\u003e \u003cp\u003eSOP 7.2.4 Notes 217\u003c\/p\u003e \u003cp\u003eSOP 7.2.5 References 217\u003c\/p\u003e \u003cp\u003eSOP 7.3 Quantification of Potentially Microcystin\/Nodularin-Producing Anabaena, Microcystis, Planktothrix, and Nodularia 218\u003cbr\u003e\u003ci\u003eAnne Rantala-Ylinen, Kaarina Sivonen, and Rainer Kurmayer\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 7.3.1 Introduction 218\u003c\/p\u003e \u003cp\u003eSOP 7.3.2 Experimental 219\u003c\/p\u003e \u003cp\u003eSOP 7.3.3 Procedure 219\u003c\/p\u003e \u003cp\u003eSOP 7.3.4 Notes 221\u003c\/p\u003e \u003cp\u003eSOP 7.3.5 References 221\u003c\/p\u003e \u003cp\u003eSOP 7.4 Relative Quantification of Microcystis or Planktothrix mcy Genotypes Using qPCR 222\u003cbr\u003e\u003ci\u003eRainer Kurmayer\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 7.4.1 Introduction 222\u003c\/p\u003e \u003cp\u003eSOP 7.4.2 Experimental 222\u003c\/p\u003e \u003cp\u003eSOP 7.4.3 Procedure 224\u003c\/p\u003e \u003cp\u003eSOP 7.4.4 Notes 225\u003c\/p\u003e \u003cp\u003eSOP 7.4.5 References 225\u003c\/p\u003e \u003cp\u003eSOP 7.5 Quantification of Transcript Amounts of mcy Genes in Planktothrix 226\u003cbr\u003e\u003ci\u003eGuntram Christiansen and Rainer Kurmayer\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 7.5.1 Introduction 226\u003c\/p\u003e \u003cp\u003eSOP 7.5.2 Experimental 226\u003c\/p\u003e \u003cp\u003eSOP 7.5.3 Procedure 227\u003c\/p\u003e \u003cp\u003eSOP 7.5.4 Notes 228\u003c\/p\u003e \u003cp\u003eSOP 7.5.5 References 228\u003c\/p\u003e \u003cp\u003eSOP 7.6 Quantification of Potentially Cylindrospermopsin-Producing Chrysosporum ovalisporum 229\u003cbr\u003e\u003ci\u003eRehab El-Shehawy and Antonio Quesada\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 7.6.1 Introduction 229\u003c\/p\u003e \u003cp\u003eSOP 7.6.2 Experimental 229\u003c\/p\u003e \u003cp\u003eSOP 7.6.3 Procedure 230\u003c\/p\u003e \u003cp\u003eSOP 7.6.4 Notes 231\u003c\/p\u003e \u003cp\u003eSOP 7.6.5 References 231\u003c\/p\u003e \u003cp\u003eSOP 7.7 qPCR Detection of the Paralytic Shellfish Toxin Biosynthesis Gene sxtB 231\u003cbr\u003e\u003ci\u003eHenna Savela\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 7.7.1 Introduction 231\u003c\/p\u003e \u003cp\u003eSOP 7.7.2 Experimental 232\u003c\/p\u003e \u003cp\u003eSOP 7.7.3 Procedure 233\u003c\/p\u003e \u003cp\u003eSOP 7.7.4 Notes 234\u003c\/p\u003e \u003cp\u003eSOP 7.7.5 References 234\u003c\/p\u003e \u003cp\u003eSOP 7.8 Application of the Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) Guidelines to Quantitative Analysis of Toxic Cyanobacteria 234\u003cbr\u003e\u003ci\u003eHenna Savela\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 7.8.1 Introduction 234\u003c\/p\u003e \u003cp\u003eSOP 7.8.2 Sampling 235\u003c\/p\u003e \u003cp\u003eSOP 7.8.3 Sample Preparation and DNA Extraction 235\u003c\/p\u003e \u003cp\u003eSOP 7.8.4 Target Information and Oligonucleotide Design 235\u003c\/p\u003e \u003cp\u003eSOP 7.8.5 qPCR Protocol 238\u003c\/p\u003e \u003cp\u003eSOP 7.8.6 qPCR Validation 239\u003c\/p\u003e \u003cp\u003eSOP 7.8.7 Data Analysis 239\u003c\/p\u003e \u003cp\u003eSOP 7.8.8 Reference 239\u003c\/p\u003e \u003cp\u003e8 DNA (Diagnostic) and cDNA Microarray 241\u003cbr\u003e\u003ci\u003eAnne Rantala-Ylinen, Kaarina Sivonen, and Annick Wilmotte\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e8.1 DNA (Diagnostic) Microarray 241\u003c\/p\u003e \u003cp\u003e8.1.1 Introduction 241\u003c\/p\u003e \u003cp\u003e8.1.2 Methodological Principles 242\u003c\/p\u003e \u003cp\u003e8.1.3 General Conclusions 243\u003c\/p\u003e \u003cp\u003e8.1.4 References 243\u003c\/p\u003e \u003cp\u003e8.2 cDNA Microarray for Cyanobacteria 244\u003cbr\u003e\u003ci\u003eHans C.P. Matthijs and J. Merijn Schuurmans\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e8.2.1 Introduction 244\u003c\/p\u003e \u003cp\u003e8.2.2 Principles of Microarray Use 244\u003c\/p\u003e \u003cp\u003e8.2.3 Considerations for Experimental Design 245\u003c\/p\u003e \u003cp\u003e8.2.4 Microarray: Practical Approach 246\u003c\/p\u003e \u003cp\u003e8.2.5 Microarray: Data Analysis 246\u003c\/p\u003e \u003cp\u003e8.2.6 References 246\u003c\/p\u003e \u003cp\u003eSOP 8.1 DNA-Chip Detection of Potential Microcystin and Nodularin Producing Cyanobacteria in Environmental Water Samples 248\u003cbr\u003e\u003ci\u003eAnne Rantala-Ylinen and Kaarina Sivonen\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 8.1.1 Introduction 248\u003c\/p\u003e \u003cp\u003eSOP 8.1.2 Experimental 249\u003c\/p\u003e \u003cp\u003eSOP 8.1.3 Procedure 250\u003c\/p\u003e \u003cp\u003eSOP 8.1.4 Notes 253\u003c\/p\u003e \u003cp\u003eSOP 8.1.5 References 253\u003c\/p\u003e \u003cp\u003eSOP 8.2 cDNA Microarrays for Cyanobacteria 254\u003cbr\u003e\u003ci\u003eJ. Merijn Schuurmans and Hans C.P. Matthijs\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 8.2.1 Introduction 254\u003c\/p\u003e \u003cp\u003eSOP 8.2.2 Experimental 254\u003c\/p\u003e \u003cp\u003eSOP 8.2.3 Procedure 256\u003c\/p\u003e \u003cp\u003eSOP 8.2.4 Notes 259\u003c\/p\u003e \u003cp\u003eSOP 8.2.5 Reference 261\u003c\/p\u003e \u003cp\u003e9 Analysis of Toxigenic Cyanobacterial Communities through Denaturing Gradient Gel Electrophoresis 263\u003cbr\u003e\u003ci\u003eIwona Jasser, Aleksandra Bukowska, Jean-Francois Humbert, Kaisa Haukka, and David P. Fewer\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e9.1 Introduction 263\u003c\/p\u003e \u003cp\u003e9.2 Main Applications of the Method 264\u003c\/p\u003e \u003cp\u003e9.3 Possible Applications 264\u003c\/p\u003e \u003cp\u003e9.4 DGGE Procedure 265\u003c\/p\u003e \u003cp\u003e9.5 General Conclusions Including Pros and Cons of the Method 267\u003c\/p\u003e \u003cp\u003e9.6 Optimization of the Method and Troubleshooting 267\u003c\/p\u003e \u003cp\u003e9.7 References 268\u003c\/p\u003e \u003cp\u003eSOP 9.1 DGGE-mcyA Conditions 270\u003cbr\u003e\u003ci\u003eAleksandra Bukowska and Iwona Jasser\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 9.1.1 Introduction 270\u003c\/p\u003e \u003cp\u003eSOP 9.1.2 Experimental 270\u003c\/p\u003e \u003cp\u003eSOP 9.1.3 Procedure 272\u003c\/p\u003e \u003cp\u003eSOP 9.1.4 Notes 275\u003c\/p\u003e \u003cp\u003eSOP 9.1.5 References 275\u003c\/p\u003e \u003cp\u003e10 Monitoring of Toxigenic Cyanobacteria Using Next-Generation Sequencing Techniques 277\u003cbr\u003e\u003ci\u003eLi Deng, Maxime Sweetlove, Stephan Blank, Dagmar Obbels, Elie Verleyen, Wim Vyverman, and Rainer Kurmayer\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e10.1 Introduction 277\u003c\/p\u003e \u003cp\u003e10.2 Specific Procedures 279\u003c\/p\u003e \u003cp\u003e10.2.1 16S rRNA Gene Amplicon Library Preparation 279\u003c\/p\u003e \u003cp\u003e10.2.2 Amplicon Purification, Quantification and Pooling 280\u003c\/p\u003e \u003cp\u003e10.2.3 Sequencing 280\u003c\/p\u003e \u003cp\u003e10.2.4 Bioinformatic Exploration of Sequencing Results 281\u003c\/p\u003e \u003cp\u003e10.2.5 General Conclusions Including Pros and Cons of the Method 281\u003c\/p\u003e \u003cp\u003e10.2.6 References 281\u003c\/p\u003e \u003cp\u003e10.3 Bioinformatic Processing of Amplicon Sequencing Datasets 283\u003cbr\u003e\u003ci\u003eMaxime Sweetlove, Dagmar Obbels, Elie Verleyen, Igor S. Pessi,\u003c\/i\u003e \u003ci\u003eAnnick Wilmotte, and Wim Vyverman\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e10.3.1 Introduction 283\u003c\/p\u003e \u003cp\u003e10.3.2 Sequencing Platforms 283\u003c\/p\u003e \u003cp\u003e10.3.3 Data Formats 284\u003c\/p\u003e \u003cp\u003e10.3.4 Error Associated with NGS Data 285\u003c\/p\u003e \u003cp\u003e10.3.5 OTU Delineation: Choosing a Similarity Threshold 286\u003c\/p\u003e \u003cp\u003e10.3.6 Conclusions 286\u003c\/p\u003e \u003cp\u003e10.4 References 286\u003c\/p\u003e \u003cp\u003eSOP 10.1 Standard Technique to Generating 16S rRNA PCR Amplicons for NGS 288\u003cbr\u003e\u003ci\u003eLi Deng, Stephan Blank, Guntram Christiansen, and Rainer Kurmayer\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 10.1.1 Introduction 288\u003c\/p\u003e \u003cp\u003eSOP 10.1.2 Experimental 288\u003c\/p\u003e \u003cp\u003eSOP 10.1.3 Procedure 289\u003c\/p\u003e \u003cp\u003eSOP 10.1.4 Notes 290\u003c\/p\u003e \u003cp\u003eSOP 10.1.5 References 290\u003c\/p\u003e \u003cp\u003eSOP 10.2 Bioinformatics Analysis for NGS Amplicon Sequencing 291\u003cbr\u003e\u003ci\u003eMaxime Sweetlove, Dagmar Obbels, Elie Verleyen, Igor S. Pessi, Annick\u003c\/i\u003e \u003ci\u003eWilmotte, and Wim Vyverman\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSOP 10.2.1 Introduction 291\u003c\/p\u003e \u003cp\u003eSOP 10.2.2 Experimental 291\u003c\/p\u003e \u003cp\u003eSOP 10.2.3 Practical Tips and Alternatives for Quality Filtering 298\u003c\/p\u003e \u003cp\u003eSOP 10.2.4 References 298\u003c\/p\u003e \u003cp\u003e11 Application of Molecular Tools in Monitoring Cyanobacteria and Their Potential Toxin Production 301\u003cbr\u003e\u003ci\u003eVitor Ramos, Cristiana Moreira, Joanna Mankiewicz-Boczek, and Vitor Vasconcelos\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e11.1 Introduction 301\u003c\/p\u003e \u003cp\u003e11.2 Possible Applications 303\u003c\/p\u003e \u003cp\u003e11.3 Checklist of Publications, Applications and Lessons from Practice 315\u003c\/p\u003e \u003cp\u003e11.3.1 Molecular-Based Studies on (Toxic) Cyanobacteria: Overview of Methods Being Used, and Generic Findings and Concerns 315\u003c\/p\u003e \u003cp\u003e11.3.2 The Need for Complementary Approaches 316\u003c\/p\u003e \u003cp\u003e11.3.3 Interpreting Results 316\u003c\/p\u003e \u003cp\u003e11.3.4 Choice of Molecular Tools for Toxigenicity Assessment 317\u003c\/p\u003e \u003cp\u003e11.3.5 Common and Possible Applications of Molecular Tools 318\u003c\/p\u003e \u003cp\u003e11.4 General Conclusions 321\u003c\/p\u003e \u003cp\u003e11.5 Acknowledgments 324\u003c\/p\u003e \u003cp\u003e11.6 References 324\u003c\/p\u003e \u003cp\u003eAppendix: Supplementary Tables 335\u003c\/p\u003e \u003cp\u003eCyanobacterial Species Cited in the Book 376\u003c\/p\u003e \u003cp\u003eGlossary 379\u003c\/p\u003e \u003cp\u003eIndex 393\u003c\/p\u003e   \u003cp\u003e\u003cb\u003e EDITED BY\u003cbr\u003e RAINER KURMAYER, PhD,\u003c\/b\u003e is an Associate Professor at the University of Innsbruck, Research Institute for Limnology, Mondsee, Austria.   \u003c\/p\u003e\u003cp\u003e\u003cb\u003e KAARINA SIVONEN, PhD,\u003c\/b\u003e is a Professor of Microbiology at the University of Helsinki, Finland.   \u003c\/p\u003e\u003cp\u003e\u003cb\u003e ANNICK WILMOTTE, PhD,\u003c\/b\u003e is a FRS-FNRS Research Associate at InBios – Center for Protein Engineering, University of Liège, Belgium.   \u003c\/p\u003e\u003cp\u003e\u003cb\u003e NICO SALMASO, PhD,\u003c\/b\u003e is Head of the Hydrobiology Unit of the Istituto Agrario di S. Michele All'Adige, Fondazione E. Mach (FEM), Trento, Italy.       \u003c\/p\u003e\u003cp\u003e In order to influence public policy regarding the detection and quantification of those organisms, it is incumbent upon scientists to raise the awareness of policy makers concerning the increased occurrence of toxigenic cyanobacteria and the threats they pose. As molecular methods can handle many samples in short time and help identify toxigenic organisms, they are reliable, cost-effective tools available for tracking toxigenic cyanobacteria worldwide. This volume arms scientists with the tools they need to track toxigenicity in surface waters and food supplies and, hopefully, to develop new techniques for managing the spread of toxic cyanobacteria.   \u003c\/p\u003e\u003cp\u003e This handbook offers the first comprehensive treatment of molecular tools for monitoring toxigenic cyanobacteria. Growing out of the findings of the landmark European Cooperation in Science and Technology Cyanobacteria project (CYANOCOST), it provides detailed, practical coverage of the full array of available molecular tools and protocols, from water sampling, nucleic acid extraction, and downstream analysis—including PCR and qPCR based methods—to genotyping (DGGE), diagnostic microarrays, and community characterization using next-gen sequencing techniques.   \u003c\/p\u003e\u003cul\u003e \u003cli\u003eOffers an overview of the latest trends in the field, while providing a foundation for understanding and applying the tools and techniques described\u003c\/li\u003e \u003cli\u003eProvides detailed coverage of the full range of molecular tools currently available, with expert guidance on the analysis and interpretation of results\u003c\/li\u003e \u003cli\u003eIncludes step-by-step guidance on standard operational procedures, including molecular tests used in environmental monitoring, with individual chapters devoted to each procedure\u003c\/li\u003e \u003cli\u003eComplements the published \u003ci\u003eHandbook of Cyanobacterial Monitoring and Cyanotoxin Analysis\u003c\/i\u003e from the CyanoCOST project\u003c\/li\u003e \u003c\/ul\u003e \u003cbr\u003e  \u003cp\u003e This handbook is an indispensable working resource for scientists, lab technicians, and water management professionals and an excellent text\/reference for graduate students and supervisors who use molecular tools. It will also be of great value to environmental health and protection officials and policy makers.\u003c\/p\u003e","brand":"Wiley","offers":[{"title":"Default Title","offer_id":47989650096357,"sku":"NP9781119332107","price":162.95,"currency_code":"USD","in_stock":false}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/1842\/7735\/files\/9781119332107.jpg?v=1761784959","url":"https:\/\/k12savings.com\/products\/molecular-tools-for-the-detection-and-quantification-of-toxigenic-cyanobacteria-isbn-9781119332107","provider":"K12savings","version":"1.0","type":"link"}