Books in the Methods in Molecular Biology series

This volume provides readers with wide-ranging coverage of CRISPR systems and their applications in various plant species. The chapters in this book discuss topics such as plant DNA repair and genome editing; analysis of CRISPR-induced mutations; multiplexed CRISPR/Cas9 systems; CRISPR-Cas12a (Cpf1) editing systems; and non-agrobacterium based CRISPR delivery systems. 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.Comprehensive and thorough, Plant Genome Editing with CRISPR Systems: Methods and Protocols is a valuable resource for any researcher interested in learning about and using CRISPR systems in plants.

This volume provides detailed descriptions on cutting-edge experimental methods to assess the multiple and complex interactions between T.

The new and updated chapters discuss the latest approaches for studying single cell RNA sequencing, whole genome sequencing data, CRISPR/Cas9-mediated gene knockout, protein interactions by a proximity ligation assay, and detection of circulating cell-free tumor DNA in lymphomas.

This book aims to provide examples of applications of atomic force microscopy (AFM) using biological samples, showing different methods for AFM sample preparation, data acquisition and processing, and avoiding technical problems. Divided into two sections, chapters guide readers through image artifacts, process and quantitatively analyze AFM images, lipid bilayers, image DNA-protein complexes, AFM cell topography, single-molecule force spectroscopy, single-molecule dynamic force spectroscopy, fluorescence methodologies, molecular recognition force spectroscopy, biomechanical characterization, AFM-based biosensor setup, and detail how to implement such an in vitro system, which can monitor cardiac electrophysiology, intracellular calcium dynamics, and single cell mechanics. 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, Atomic Force Microscopy: Methods and Protocols is useful for researchers at different stages, from newcomers to experienced users, interested in new AFM applications.

This volume provides researchers with up-to-date protocols and takes aclose look at current research and promising applications.

This volume covers arange of methods used in plant cytogenetics, beginning with basic analysis ofchromosomes and visualizing gene locations, to manipulating and dissectingchromosomes, and then focusing on less understood features of chromosomes suchas recombination initiation sites and epigenomic marks.

This book provides coverage, methodology, and laboratory protocols on the more essential aspects of protein tyrosine phosphatase (PTP) function and regulation, including the use of standardized in vitro functional assays, suitable cell systems, and animal and microorganism models.

This volume highlights the role of proteostasis in human health and associated disease model systems, reflecting its rising importance which has led to the development of new technologies to obtain insight into underling protein mechanistic events.

This volume is the result of an explosion ofmolecular-based research on Cilia, which began with the discovery of theuniversality of intraflagellar transport (IFT) and ciliary genomics/proteomics.The chapters in this book cover topics such as: high resolution imaging andfunctional characterization of sensory and primary cilia in mammalian cells andzebrafish, methods to study ciliary-mediated chemoresponse in Paramecium,and methods to study centrosomes and cilia in C. elegans and Drosophila.Written in the highly successful Methods in Molecular Biology series format,chapters include introduction to their respective topics, lists of thenecessary materials and reagents, step-by-step, readily reproducible laboratoryprotocols, and tips on troubleshooting and avoiding known pitfalls.Practical and cutting-edge, Cilia:Methods and Protocols is broad and covers motile, sensory, and primarycells. It is a valuable resource to anyone interested in entering the field ofciliary biology using model organisms, including flagellate algae, ciliates,planaria, nematodes, insects, zebrafish, and mammalian cells.

This volume provides an up-to-date compilationof current methodological approaches utilized for the exploration of nucleolarstructure and function. Chapters covera diversity of protocols that include imaging of the nucleolus, analysis ofribosomal RNA transcription and processing, and genomics and proteomics of thenucleolus.

This volume presents methods and techniques to studyoogenesis in a broad range of organisms, from plants to mammals. Oogenesis: Methods and Protocols guidesreaders through protocols on models ofdevelopmental biology, oogenesis in plants, worms, fruit flies,mosquitos, butterflies, starfish, zebrafish, frog, chicken and mouse. Written in the highly successful Methodsin Molecular Biology series format, chapters include introductions to theirrespective topics, lists of the necessary materials and reagents, step-by-step,readily reproducible laboratory protocols, and tips on troubleshooting andavoiding known pitfalls.Authoritative and cutting-edge, Oogenesis:Methods and Protocols aims to ensure successful results in the furtherstudy of this vital field.

This book covers core and emerging in vitro and in vivo protocols used to study how various components of the tumor microenvironment are established and subsequently interact with tumor cells to facilitate carcinogenesis.

This volume presents relevantbackground information to understanding the molecular basis governingunconventional protein secretion (UPS), and in particular explores the latesttechniques and protocols that have been successfully applied for the study ofthis topic. Detailed chapters include an overview of conventional andunconventional secretory pathways along with multidisciplinary approaches andmethods used for UPS analysis in different organisms. Written in the highly successful Methods inMolecular Biology series format, chapters include introductions to theirrespective topics, lists of the necessary materials and reagents, step-by-step,readily reproducible laboratory protocols, and tips on troubleshooting andavoiding known pitfalls. Authoritative and cutting-edge, UnconventionalProtein Secretion: Methods and Protocols will be usefulfor all interested in the secretory pathway field as well as applications incell biology, cell development, biomedical research, and healthcare.

Chapters address the three major areas of study: provoking regeneration by inducing damage to muscle, analyzing theprogenitor cells of skeletal muscle, and quantifying overall muscle function.

This volume discusses experimental brain injury models thatcontain valuable information carefully chosen to widen the researchers¿ horizonabout neurotrauma. Injury Models of Central Nervous System: Methods and Protocols contains relevant experimental design approaches that have beenadapted and made ready for application in laboratory settings. For easiernavigation, the chapters are categorized into 6 parts: Introduction, GeneralConsideration in Using Animal Laboratory in CNS Injury Research, Classical TBIModels and Their Link with Pathophysiological Features of CBS Injury ¿ Models,Special Topics in CNS Trauma: Comorbid Conditions in CNS Injury, OutcomeMeasures in Brain Injury Models, and Future Directions. Written in the highly successful Methods inMolecular Biology series format, chapters include introductions to theirrespective topics, lists of the necessary materials and reagents, step-by-step,readily reproducible laboratory protocols, and tips on troubleshooting andavoiding known pitfalls.Practical and thorough, Injury Models of Central Nervous Systems: Methods and Protocols, is a very useful reference towards theprogress of this discipline.

This volume highlights the molecular and cellular methods used in studying Chronic Myeloid Leukimia (CML) pathogenesis and stem cell biology. 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, Chronic Myeloid Leukemia: Methods and Protocols aims to ensure successful results in the further study of this vital field.

This second edition provides updated and new chapters on selected genetic, molecular, biochemical, and cell biological techniques. Chapter's guide readers through methods and principles on primordial germ cells and germline stem cells, however many of these principles can be applied to different types of adult stem cells.

This volume provides a comprehensive overview of the experimental and computational methodologies used to study the function of long non-coding RNA (ncRNAs) expressed from enhancers. Chapter detail both wet-lab and dry-lab techniques and annotating long ncRNAs and exploring transcription by assessing where transcription starts and generally how it occurs.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, Enhancer RNAs: Methods and Protocols aims to ensure successful results in this rapidly developing field.

High-throughput RNAi screening remains one of the most widely used technologies to perform target identification and validation studies in an unbiased manner. These assays are equally important for research and development across academic, biotech, and pharmaceutical industries. The success of these screening efforts is dependent on robust methodologies to perform these screens. In High-Throughput RNAi Screening: Methods and Protocols, expert researchers in the field share protocols and methods for performing high-throughput RNAi (HT-RNAi) screens. These include the use of various RNAi platforms and delivery methods in mammalian and non-mammalian systems, whole organism and cell models, and various applications, such as drug sensitizer identification. Finally, the book examines the latest advancements in the fields of assay development, library screening, data analysis, and hit selection. 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 thorough, High-Throughput RNAi Screening: Methods and Protocols provides a comprehensive source of protocols and other necessary information to make robust and successful assays possible for all who wish to apply HT-RNAi in their research.

This volume presents authoritative and cutting-edge methods and protocols focusing on three tool boxes covering the increasingly diverse methodologies used to image selected proteins and to investigate their function by light and electron microscopy.

This second edition provides 21 new chapters on methods used in laboratories for investigating the physiology and molecular genetics of the pathogen Clostridium difficile.

This volume provides a unique forum to review cell microencapsulation in a broad sense by exploring various cell types that have been encapsulated for different purposes, different approaches and devices used for microencapsulation, the biomaterials used in cell microencapsulation, the challenges to the technology, and the current status of its application in different clinical situations. This book is divided in five sections: Section I is an introductory part that discusses historical developments of the technology and its current challenges, as well as the various applications of cell microencapsulation; Section II discusses the main approaches and devices currently used in cell microencapsulation; Section III presents an overview of the various polymeric materials currently in use for cell microencapsulation and the enabling technologies to either monitor or enhance encapsulated cell function; Section IV gives specific examples of the methods used to encapsulate various cell types; and Section V provides an overview of the different clinical situations in which cell microencapsulation has been applied. 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.Thorough and practical, Cell Microencapsulation: Methods and Protocols is a valuable reference for researchers, engineers, clinicians, and other healthcare professionals, as well as food technologists who will find detailed descriptions of methods for the microencapsulation of specific cell types and their current of potential clinical and industrial applications. This volume also includes detailed information about the design and manufacture of different devices including large-scale production devices for use in cell microencapsulation.

Now poised to evaluate the potential in modulating Wnt signaling for therapeutic agendas in cancer, wound healing, and degenerative disease, this book collects protocols from an exciting area of study.

Practical and cutting-edge, Plant Synthetic Promoters: Methods and Protocols covers all steps required from the identification of cis-regulatory elements, over synthetic promoter design, to the experimental analysis of synthetic promoter function.

This thorough volume explores predicting one-dimensional functional properties, functional sites in particular, from protein sequences, an area which is getting more and more attention. Beginning with secondary structure prediction based on sequence only, the book continues by exploring secondary structure prediction based on evolution information, prediction of solvent accessible surface areas and backbone torsion angles, model building, global structural properties, functional properties, as well as visualizing interior and protruding regions in proteins. Written for the highly successful Methods in Molecular Biology series, the chapters include the kind of detail and implementation advice to ensure success in the laboratory. Practical and authoritative, Prediction of Protein Secondary Structure serves as a vital guide to numerous state-of-the-art techniques that are useful for computational and experimental biologists.

This volume explores a broad range of different genotyping techniques. Genotyping: Methods and Protocols consists of chapters that cover numerous topics such as: an overview of multiplexed microsatellite analysis; High Resolution Melt analysis and TaqMan-based assays; in situ analysis of variants in single RNA molecules; the MassARRAY system and Molecular Inversion Probes; Pulsed Field Gel Electrophoresis, Paralogue Ratio Test, and Multiplex Ligation-Dependent Probe Amplification; long-range PCR combined with PacBio sequencing; Targeted Locus Amplification; Multilocus Sequence Typing and rapid SNP detection with pyrosequencing; and genotyping-by-sequencing for plant analysis. Finally, the volume concludes with a summary of pertinent points to describe genetic variation. 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.Thorough and practical, Genotyping: Methods and Protocols is a valuable resource for anyone interested in learning more about the diverse field of genotyping.

This collection seeks to elucidate the practical methods necessary for successful adjuvant development, with a particular focus on the synthesis, formulation, manufacturing, and characterization aspects involved. Beginning with an overview and a case study, the book then delves into in silico design, chemical synthesis, biosynthesis, and/or purification from natural raw materials of specific adjuvant molecules, adjuvant formulation approaches, the analytical characterization of adjuvant formulations and adjuvant-containing vaccines, as well as the biological characterization of vaccine adjuvant activity, including in vitro and in vivo approaches, to measure innate and adaptive immune responses. Written in the highly successful Methods in Molecular Biology 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.Practical and authoritative, Vaccine Adjuvants: Methods and Protocols aims to facilitate vaccine adjuvant product development efforts, making them more accessible, manufacturable, and better characterized.

This volume aims to present a representative cross-section of modern experimental approaches relevant to Plant Hormone Biology, ranging from relatively simple physiological to highly sophisticated methods. Chapters describe physiological, developmental, microscopy-based techniques, measure hormone contents, and heterologous systems. 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, Plant Hormones:Methods and Protocols, Third Edition aims to provide researchers with useful methods to advance their research.

This volume describes a variety of protocols that will allow the readers to study different aspects of transcriptional and posttranscriptional gene expression regulation in eukaryotic cells.

This detailed volume presents protocols for advancing the utility of nanotechnology in cancer research toward improving our understanding of cancer biology, prevention, diagnosis, and therapy. There are continuous new discoveries in the field of nanotechnology, thus creating new imaging systems or therapies, and this book focuses on how to employ certain discoveries for studying cancer by presenting principles along with techniques to allow for the transformation of any new discoveries in the field into cancer-studying tools with the hope of bringing in the involvement of biomedical scientists who can enhance the speed of discoveries toward cancer diagnosis and therapy. 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 motivating, Cancer Nanotechnology: Methods and Protocols serves as an ideal resource for biomedical scientists interested in the potential of this field as well as for physical scientists and engineers interested in employing nanotechnology in cancer diagnosis and therapy.