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This volume details the fundamentals of the CRISPR-Cas system, and its protocols illustrate advances in CRISPR-Cas techniques for efficient genome editing. Introductory chapters provide a wide horizon of CRISPR/Cas-based methods and applications. Additional chapters guide readers through HDR-mediated editing, sgRNA design, the step-by-step procedure of multiplex adenine base editing experiments in rice, generating mutants for rice, wheat, Brachypodium, Barley, Flax, and Phytophthora, visual screening of mutants, gene deletion (knock-out), tagging (knock-in) in mammalian cells, the cloning-free (DNA-free) technique, cell-penetrating peptides, generating a genome-edited banana, and nuclear genome editing of Chlamydomonas employing CRISPR-Cpf1 combined with a single-stranded DNA (ssODN) repair template. Authoritative and cutting-edge, CRISPR-Cas Methods aims to assist researchers who are new to the field and are aiming to learn how best to adopt this technology for a particular organism.
This volume details protocols encompassing different aspects of computer aided design and manufacturing of 3D scaffolds and biofabricated constructs for tissue engineering applications.
This volume details protocols encompassing different aspects of computer aided design and manufacturing of 3D scaffolds and biofabricated constructs for tissue engineering applications.
This second edition provides techniques used to study of the underlying transcriptional programs of stem cells that promote self-renewal and differentiation. Chapters detail next-generation sequencing technologies, data analysis, protocols on analysis and visualization of single-cell RNA-Seq data, analysis of 3D chromatin architecture, interpretation of large-scale interaction networks, transcriptional networks in embryonic and adult stem cells, derivation of stem cells, and transcriptional programs that promote reprogramming, transdifferentiation, and cancer formation. 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, Stem Cell Transcriptional Network: Methods and Protocols, Second Edition aims to provide a key resource for biologists seeking to interrogate these vital networks.
This volume looks at the latest techniques used to perform comparative structure analyses, and predict and evaluate protein-ligand interactions. The chapters in this book cover tools and servers such as LiteMol; Bio3D-Web; DALI; CATH; HoTMuSiC, a contact-base protein structure analysis tool known as CAD-Score; PyDockSaxs and HADDOCK; CombDock and DockStar; the BioMagResBank database; as well as BME and CoNSEnsX+. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, step-by-step, readily reproducible computational protocols, and tips on troubleshooting and avoiding known pitfalls.Cutting-edge and comprehensive, Structural Bioinformatics: Methods and Protocols is a practical guide for researchers to learn more about the aforementioned tools to further enhance their studies in the growing field of structural bioinformatics.Chapter 13 is available open access under a CC-BY 4.0 license via link.springer.com.
This volume provides current and new advanced methods and protocols to study T cells. Chapters guide readers through T cell diversity using mass cytometry, analyzing T cells from single cell level, CRISPR/Cas9 techniques to study the T cell activation, techniques to study subsets of TcellΓÇÖs, procedures to study artificial antigen presentosomes for T cell activation, techniques to study the T cell development, two-photon microscopy, and MAIT cells. 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, T-Cell Receptor Signaling: Methods and Protocols aims to provide a wide range of approaches and be an invaluable resource for present and future generations of T cell researchers.
This volume details methods and protocols necessary to further the study of insect immunity. Chapters guide readers through up-to-date genomic and transcriptomic approaches, insect samples for proteomic analysis, hemocytes in Drosophila, cellular response in Lepidoptera, insect AMPs, manipulate the composition of mosquito microbiota, viral infections in insects, infections by entomopathogenic nematodes, immune response following oral infections, and protocols to to monitor the effect of septic infections with human pathogens using B. mori as a model. 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, Immunity in Insects aims to ensure successful results in the further study of this vital field.
This book provides a comprehensive collection of classic and cutting-edge methodologies as well as bioinformatics and genome-editing approaches that are used to quantify immune mediators and analyze their function and biological activity in cancer cells and tissues.
This book provides a comprehensive guide to scientists, engineers, and students that employ metabolomics in their work, with an emphasis on the understanding and interpretation of the data. Chapters guide readers through common tools for data processing, using database resources, major techniques in data analysis, and integration with other data types and specific scientific domains. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, practical guidance of methods and techniques, useful web supplements, and connect the steps from experimental metabolomics to scientific discoveries.Authoritative and cutting-edge, Computational Methods and Data Analysis for Metabolomics to ensure successful results in the further study of this vital field.
This fully revised book includes new and improved protocols to analyze toxicity at the DNA, RNA, and protein levels using genomic, epigenomic, transcriptomic, and proteomic approaches. Sections of the book address key subjects such as bioinformatics and biostatistics, toxicant-induced lung inflammation and tumors, analysis of protein and gene expression and silencing, gene promoter methylation, detection of chromosomal and genome-wide damage, analysis of DNA-adducts and surrogate gene mutations, detection and characterization of cancer gene mutation, and many other areas of study. 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. Comprehensive and up-to-date, Molecular Toxicology Protocols, Third Edition will prove to play an important role in generating a better understanding of the field of toxicology and its sub-disciplines.
Animal Cell Biotechnology: Methods and Protocols, Fourth Edition constitutes a comprehensive manual of state-of-the-art techniques for setting up mammalian cell lines and media for development of biopharmaceuticals, and optimizing critical parameters for cell culture considering the whole cascade from the lab to the final production.
This volume looks at the latest technologies and methods used by researchers to study mechanisms that control the aspects of plant stem cell signaling events.
This volume provides comprehensive methods from expert scientists working in the Chimeric Antigen Receptor T Cell (CAR-T Cell) field. Chapters guide readers through the state-of-art of CAR-T cell technology, CAR design and vector production, CAR-T cell generation and manufacturing, CAR-T cell characterization, and quality control.
This volume details state-of-art eQTL analysis, where interdisciplinary researchers are provided both theoretical and practical guidance to eQTL analysis and interpretation.
This book highlights many fundamental aspects of optical fiber transmission engineering while also focusing on current state of the art applications and working examples of digital coherent optical communications. Major engineering themes are reviewed and analyzed in this book, including spectral and time-domain characteristics of multi-level pseudo-random PAM signals, optical QAM and SSB complex modulations and impulse response engineering of linear amplifiers used in next-generation Gbaud transmission systems. This book is balanced between theoretical and numerical simulation approaches, showing numerous working examples developed in Matlab.Presents an in-depth analysis of pseudo-random multi-level signals and high-order complex modulations to support coherent terabit transmission systems;Provides a unified approach to challenging engineering issues encountered in the design of Giga-baud coherent optical transmission systems using high-order complex modulation formats;Reviews engineering themes and provides in-depth analysis, modeling and quantitative examples and solutions of state of the art and future applications.
This book of protocols is devoted to the yellow coenzymes derived from riboflavin or vitamin B2 and to the hundreds of enzymes whose functionality depends on them, and represents a compendium of techniques for working with flavoproteins or with the wide spectrum of functions that flavoproteins can drive in the cells. Starting with Rf production in microorganisms and the chemical, optical, and redox properties of these fascinating molecules and moving along to the variety and the peculiarity of some single flavoenzymes, the volume explores the complexity of functions and distribution of these molecules in the cell. 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, Flavin and Flavoproteins: Methods and Protocols serves as an ideal guide for protein chemists interested in purifying and characterizing flavoproteins, as well as microbiologists, physiologists, and clinicians, who wish to further study problems connected with flavoproteins.
This book covers two major classes of mixed effects models, linear mixed models and generalized linear mixed models. It presents an up-to-date account of theory and methods in analysis of these models as well as their applications in various fields.
This book highlights the importance of the choroid plexus, which forms the blood-cerebrospinal fluid barrier and is the site of the major production of the cerebrospinal fluid (CSF).
This second edition offers 88 chapters divided among three volumes providing the most comprehensive source of know-how in the wide-ranging field of Mitochondrial Medicine. Volume II guides readers through chapters on mitochondrial dysfunction, functional¿ mitochondria, mitochondrial retrograde, mitochondrial dNTP pool quantification, mitochondrial ADP-ribosylation, blue-native gel approach, 3D optical cryo-imaging method, mitochondrial ATP and ROS production, protocol for untargeted metabolomic analysis, and methods for analysis of nitrotyrosine-containing proteins. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, application details for both the expert and non-expert reader, and tips on troubleshooting and avoiding known pitfalls. Authoritative and accessible, Mitochondrial Medicine, Second Edition, Volume 2: Assessing Mitochondria aims to be acomprehensive source of know-how in the wide-ranging field of Mitochondrial Medicine.
Though the authors have no definitive solution to the problem, they offer this contribution in an attempt to define the problem as they see it, and to sketch out several obvious attempts that have been suggested to solve the problem and which seem to have failed.
This comprehensive laboratory manual describes the various protocols involved in Actinobacterial research. The content is divided into fifteen major sections, each of which is further divided into sub-sections describing the respective aim, principles, materials & methods, protocol, expected results and diagrams.
his work over the past two decades has set the stage for medical innovation through nanotechnology. Written by world-renowned experts in the field, this book pays tribute to Dr. Mauro Ferrari's vast contribution to the field of nanotechnology and nanomedicine.
The book provides the readers of various discipline an easy understanding of the latest biophysical techniques pertaining to microbiology. Biofilm associated chronic infection is a major health problem and a serious concern to doctors, scientists and other health workers as it develops antibiotic and multi-drug resistance. This book describes various protocols utilized in the detection of the biofilm. The book has been divided into six sub sections which provides pertinent information about the various biophysical techniques and instruments that are used for detecting and analyzing the biofilm formation upon biotic and abiotic surfaces. The readers will be able to identify the techniques that can best cater information to solve the problem at hand. This book attempts to compile the latest information on the recent advances in the various functional aspects of microbial biofilms, their pathogenesis, present day treatments as well as detection strategies.This book is meant forresearchers in the field of microbiology and interested in understanding microbial pathogenesis, quorum sensing and biofilm formation.
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