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This volume provides readers with a collection of the latest protocols used to study plant genome editing and trait engineering. The chapters in this book are organized into five parts: genome engineering systems; machinery design and validation; delivery tools; generation and analysis of engineering materials; and crop genome engineering applications. The chapters cover topics such as methods of applying the popular CRISPR-Cas9 or CRISPR-Cas12 systems for editing genomes in different crop species, the use of small synthetic plastome for potato genome engineering, and the use of CRISPR-Cas9 for algal cell genome engineering. 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.Cutting-edge and comprehensive, Plant Genome Engineering: Methods and Protocols is a valuable tool for researchers interested in learning more about this developing and important field.

This new collection features the most up-to-date essential protocols that are currently being used to study the immune synapse. Beginning with methods for making biophysical measurements, the volume continues by covering the cell biology of synapses, methods for advanced substrate engineering, mechanobiology topics, new technologies to describe and manipulate synaptic components, as well as methods related to sites of action and immunotherapy. 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and fully updated, The Immune Synapse: Methods and Protocols, Second Edition serves as an ideal practical guide for researchers working in this dynamic field. Chapters 5, 11,18, 27, 30, and 32 are available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

This volume provides cutting-edge techniques to further the study chromatin biology. Chapters include both novel and well-established methods for the analysis of DNA-associated proteins, DNA methylation, three-dimensional chromatin interactions, deep sequencing-based tools, and data analysis pipelines. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, provides details of the necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and describes step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Chromatin: Methods and Protocols aims to further the understanding of how modified DNA and associated proteins affect the transcriptional output of the genome.Chapter Genome-wide mapping and microscopy visualization of protein-DNA interactions by pA-DamID [Chapter 12] is available open access undera Creative Commons Attribution 4.0 International License via link.springer.com.

This detailed book builds on the foundation of lymphatic protocols with tools to aid in the interpretation of large datasets created by angiogenesis research. For example, this includes protocols to explore how transcriptional phenotyping can be achieved, both in terms of exploring tissue-specific heterogeneity to the use of database mining of existing datasets. Similarly, bioinformatic techniques run alongside protocols for transcript analysis, cell biology, pathology, as well as physiological and developmental angiogenesis models. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Angiogenesis: Methods and Protocols provides a meaningful compendium of practical approaches to angiogenesis and endothelial cell biology research.

This volume serves to aid researchers working in the recombinant protein production field by describing a wide number of protocols and examples. Chapters describe recombinant protein production in different expression systems, prokaryotic and eukaryotic expression systems, purification protocols, characterization of insoluble proteins and a general overview of interesting applications of insoluble proteins. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Insoluble Proteins: Methods and Protocols, Second Edition aims to be a useful practical guide to researches to help further their study in this field.

This detailed volume compiles key methods and techniques used to establish some of the structural and functional aspects of Z-form nucleic acids. Beginning with chapters on characterizing Z-DNA, the book continues by exploring structure determination techniques, gene- and genome-targeting methods and genome-wide approaches, Z-RNA study, 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Z-DNA: Methods and Protocols serves as an ideal guide for students and researchers curious about this unique Z nucleic acid as an area of investigation, aiming to help the field grow, flourish, and unravel the hidden and novel roles of Z. Chapter 1 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.This is an open access book.

This volume explores the latest techniques in inner ear development, analysis of its sensory cells, and characterization and manipulation of the central auditory and vestibular pathways. The chapters in this book cover topics such as dissection and imaging of the cochlea; behavioral evaluation of animal models of diseases like tinnitus; hair cell function and regeneration; and recent advances in sequencing technology. In the Neuromethods series style, chapters include the kind of detail and key advice from the specialists needed to get successful results in your laboratory. Cutting-edge and comprehensive, Developmental, Physiological, and Function Neurobiology of the Inner Ear is a valuable resource for scientists and researchers interested in learning more about this developing field.

This detailed volume explores perspectives and methods using cell-free expression (CFE) to enable next-generation synthetic biology applications. The first section focuses on tools for CFE systems, including a primer on DNA handling and reproducibility, as well as methods for cell extract preparation from diverse organisms and enabling high-throughput cell-free experimentation. The second section provides an array of applications for CFE systems, such as metabolic engineering, membrane-based and encapsulated CFE, cell-free sensing and detection, and educational kits. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Cell¿Free Gene Expression: Methods and Protocols serves as an idealguide for researchers seeking technical methods to current aspects of CFE and related applications.

This volume details methods and protocols covering multiple aspects of Medulloblastoma. Divided into four parts, chapters guide readers through nucleic acids detection and analysis, cell-based analysis methodologies, and applications of patient-information on designing better experimental strategies for future drug development efforts in Medulloblastoma. 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, Medulloblastoma: Methods and Protocols aims to deliver a clear-cut and standardized set of protocols to a broad scientific community.

This book details the development of methods and models to study the HIV-1 viral reservoir with the ultimate goal of achieving a functional cure of HIV infection. Chapters are divided into six parts covering cell lines, in vitro and ex vivo primary cell models of persistent infection, in vitro and ex vivo tissue-derived models, infected animal models human immune cells, methods of detection and analysis of the reservoir, and current approaches to achieve either a functional cure or cART-free long-term remission. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, HIV Reservoirs: Methods and Protocols provides a comprehensive, updated collection of state-of-art methodologies and models to tackle the HIV-1 viral reservoir.

The volume presents a small selection of state-of-the-art approaches for studying transposable elements(TE). Chapters guide readers through HTS-based approaches, bioinformatic tools, methods to studyTE protein complexes, and the functional impact on the host. 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 cutting-edge, Transposable Elements: Methods and Protocols aims to be a useful practical guide to researches to help further their study in this field. 

This detailed volume explores the most recent methodologies and applications of stable isotope labeling by amino acids in cell culture (SILAC). With the development of new technologies in the field of mass spectrometry and bioinformatics, SILAC-based methods have become the first tool of choice in quantitative proteomics either on its own or in combination with other more recently developed approaches, which is elucidated in the chapters of this book. 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, SILAC: Methods and Protocols serves as an ideal guide for researchers working in the area of functional proteomics and other aspects of protein science.

This detailed book brings together a collection of methodologies, from the most basic to the more complex, that provides researchers with a platform they can use to embark on a cartilage research career. To aid in the search for novel therapies for cartilage regeneration, this volume addresses 3D cartilage models, challenges associated with RNA and protein extraction, imaging, gene transfer, as well as stable differentiation and variations in cell phenotype from different tissue origins. 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Cartilage Tissue Engineering serves as an ideal guide for researchers working to advance the vital study of cartilage biology and repair.

This book covers various state-of-the-art techniques regarding the associations between transcription factors (TFs) and genes, with a focus on providing methodological and practical references for researchers. The contents cover diverse protocols and summaries of TFs including screening of TF-DNA interactions, detection of open chromatin regions, identification of epigenetic regulations, engineering TFs with genome editing tools, detection of transcriptional activities, computational analysis of TF networks, functions and druggabilities of TFs in biomedical research, and much more. Written for the highly successful Methods in Molecular Biology series, chapters feature the kind of detailed implementation advice from the experts to ensure successful research results. Authoritative and cutting-edge, Transcription Factor Regulatory Networks aims to benefit readers who are interested in using state-of-the-art techniques to study TFs and their myriad effects in cellular life.

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.

 This second edition compiles state-of-art protocols to study the molecular mechanisms that determine proteome composition and contribute to maintaining protein homeostasis. Chapters describe methods that range from the in vitro analysis of enzyme cascades to real-time in vivo studies. 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 cutting-edge, Plant Proteostasis: Methods and Protocols, Second Edition aims to be a useful practical guide to researches to help further their study in this field.