Download and read online Crispr Cas A Laboratory Manual in PDF and EPUB The development of CRISPR-Cas technology is revolutionizing biology. Based on machinery bacteria use to target foreign DNA, these powerful techniques allow investigators to edit DNA sequences and modulate gene expression more rapidly and accurately than ever before. Featuring contributions from leading figures in the CRISPR-Cas field, this laboratory manual presents a state-of-the-art guide to the technology. It includes step-by-step protocols for applying CRISPR-Cas-based techniques in various systems, including yeast, zebrafish, Drosophila, mice, and cultured cells (e.g., human pluripotent stem cells). The contributors cover web-based tools and approaches for designing guide RNAs that precisely target genes of interest, methods for preparing and delivering CRISPR-Cas reagents into cells, and ways to screen for cells that harbor the desired genetic changes. Strategies for optimizing CRISPR-Cas in each system-especially for minimizing off-target effects-are also provided. Authors also describe other applications of the CRISPR-Cas system, including its use for regulating genome activation and repression, and discuss the development of next-generation CRISPR-Cas tools. The book is thus an essential laboratory resource for all cell, molecular, and developmental biologists, as well as biochemists, geneticists, and all who seek to expand their biotechnology toolkits.
Download and read online RNA in PDF and EPUB Almost all molecular and cellular biology laboratories now handle RNA and this manual is an authoritative source of information and protocols for this purpose, from the basic to the advanced. Required reading for every research laboratory in the life sciences.
Download and read online CRISPR Cas Systems in PDF and EPUB CRISPR/Cas is a recently described defense system that protects bacteria and archaea against invasion by mobile genetic elements such as viruses and plasmids. A wide spectrum of distinct CRISPR/Cas systems has been identified in at least half of the available prokaryotic genomes. On-going structural and functional analyses have resulted in a far greater insight into the functions and possible applications of these systems, although many secrets remain to be discovered. In this book, experts summarize the state of the art in this exciting field.
Download and read online Molecular Cloning in PDF and EPUB The first two editions of this manual have been mainstays of molecular biology for nearly twenty years, with an unrivalled reputation for reliability, accuracy, and clarity. In this new edition, authors Joseph Sambrook and David Russell have completely updated the book, revising every protocol and adding a mass of new material, to broaden its scope and maintain its unbeatable value for studies in genetics, molecular cell biology, developmental biology, microbiology, neuroscience, and immunology. Handsomely redesigned and presented in new bindings of proven durability, this three–volume work is essential for everyone using today’s biomolecular techniques. The opening chapters describe essential techniques, some well–established, some new, that are used every day in the best laboratories for isolating, analyzing and cloning DNA molecules, both large and small. These are followed by chapters on cDNA cloning and exon trapping, amplification of DNA, generation and use of nucleic acid probes, mutagenesis, and DNA sequencing. The concluding chapters deal with methods to screen expression libraries, express cloned genes in both prokaryotes and eukaryotic cells, analyze transcripts and proteins, and detect protein–protein interactions. The Appendix is a compendium of reagents, vectors, media, technical suppliers, kits, electronic resources and other essential information. As in earlier editions, this is the only manual that explains how to achieve success in cloning and provides a wealth of information about why techniques work, how they were first developed, and how they have evolved.
Download and read online CRISPR in PDF and EPUB This volume presents a list of cutting-edge protocols for the study of CRISPR-Cas defense systems and their applications at the genomic, genetic, biochemical and structural levels. CRISPR: Methods and Protocols guides readers through techniques that have been developed specifically for the analysis of CRISPR-Cas and techniques adapted from standard protocols of DNA, RNA and protein 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, CRISPR: Methods and Protocols provides a broad list of tools and techniques to study the interdisciplinary aspects of the prokaryotic CRISPR-Cas defense systems.
Download and read online The Use of CRISPR cas9 ZFNs TALENs in Generating Site Specific Genome Alterations in PDF and EPUB This new volume of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers recent research and methods development for changing the DNA sequence within the genomes of cells and organisms. Focusing on enzymes that generate double-strand breaks in DNA, the chapters describe use of molecular tools to introduce or delete genetic information at specific sites in the genomes of animal, plant and bacterial cells. Continues the legacy of this premier serial with quality chapters authored by leaders in the field Covers research methods in biomineralization science Contains sections on such topics as genome editing, genome engineering, CRISPR, Cas9, TALEN and zinc finger nuclease
Download and read online Targeted Genome Editing Using Site Specific Nucleases in PDF and EPUB This book serves as an introduction to targeted genome editing, beginning with the background of this rapidly developing field and methods for generation of engineered nucleases. Applications of genome editing tools are then described in detail, in iPS cells and diverse organisms such as mice, rats, marine invertebrates, fish, frogs, and plants. Tools that are mentioned include zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and CRISPR/Cas9, all of which have received much attention in recent years as breakthrough technologies. Genome editing with engineered nucleases allows us to precisely change the target genome of living cells and is a powerful way to control functional genes. It is feasible in almost all organisms ranging from bacteria to plants and animals, as well as in cultured cells such as ES and iPS cells. Various genome modifications have proven successful, including gene knockout and knock-in experiments with targeting vectors and chromosomal editing. Genome editing technologies hold great promise for the future, for example in biomedical research, clinical medicine, and generation of crops and livestock with desirable traits. A wide range of readers will find this book interesting, and with its focus on applications in a variety of organisms and cells, the book will be valuable for life scientists in all fields.
Download and read online The Condensed Protocols from Molecular Cloning a Laboratory Manual in PDF and EPUB The Condensed Protocols From Molecular Cloning: A Laboratory Manualis a singleâ€“volume adaptation of the threeâ€“volume third edition of Molecular Cloning: A Laboratory Manual.This condensed book contains only the stepâ€“byâ€“step portions of the protocols, accompanied by selected appendices from the world's bestâ€“selling manual of molecular biology techniques. Each protocol is crossâ€“referenced to the appropriate pages in the original manual. This affordable companion volume, designed for bench use, offers individual investigators the opportunity to have their own personal collection of short protocols from the essential Molecular Cloning.
Download and read online Live Cell Imaging in PDF and EPUB Recent advances in imaging technology reveal, in real time and great detail, critical changes in living cells and organisms. This manual is a compendium of emerging techniques, organized into two parts: specific methods such as fluorescent labeling, and delivery and detection of labeled molecules in cells; and experimental approaches ranging from the detection of single molecules to the study of dynamic processes in organelles, organs, and whole animals. Although presented primarily as a laboratory manual, the book includes introductory and background material and could be used as a textbook in advanced courses. It also includes a DVD containing movies of living cells in action, created by investigators using the imaging techniques discussed in the book. The editors, David Spector and Robert Goldman, whose previous book was Cells: A Laboratory Manual,are highly respected investigators who have taught microscopy courses at Cold Spring Harbor Laboratory, the Marine Biology Laboratory at Woods Hole, and Northwestern University.
Download and read online Forensic DNA Biology in PDF and EPUB A collection of forensic DNA typing laboratory experiments designed for academic and training courses at the collegiate level.
Download and read online Calcium Techniques in PDF and EPUB "This laboratory manual provides step-by-step protocols for studying many facets of Ca2+ signaling, as well as background information on the principles and applications of the techniques. Contributors discuss how to use fluorescent, luminescent, and genetically encoded Ca2+ probes in conjunction with state-of-the-art imaging modalities to characterize Ca2+ signals. Electrophysiological measurements of Ca2+ channel activity are described, as are radioactive Ca2+ flux assays and methods to investigate signaling mediated by specific Ca2+-mobilizing messengers (IP3, cADPR, and NAADP). Techniques to modulate and suppress intra- and intercellular signals are also provided. Each protocol is complete with a list of required materials, detailed recipes for media and reagents, and troubleshooting advice"--
Download and read online Manipulating the Mouse Embryo in PDF and EPUB This fourth edition of "The Mouse Manual¨is once again the definitive reference source on mouse development, transgenesis techniques, and molecular biology. Its preeminent authors Richard Behringer, Marina Gertsenstein, Kristina Nagy, and Andras Nagy have reorganized and updated this edition to include new information and protocols on: · generation of induced pluripotent stem cells · RNA microinjections · lentiviral microinjections and infection · assisted reproduction techniques for sperm and embryo cryopreservation · isolation, generation, and transplantation of spermatogonial stem cell lines · in utero electroporation of gene constructs into postimplantation embryos · vibratome sectioning of live and fixed tissues for imaging thick tissue sections · whole-mount fluorescent staining methods for three-dimensional visualization Recombinant DNA techniques and methods for studies of mouse embryonic development have been updated from previous editions, as has the wealth of information on mouse laboratory strains, mouse housing and breeding, surgical procedures, assisted reproduction, handling of embryos, and micromanipulation setups. The first edition of this classic work appeared more than 20 years ago, with authors that included Brigid Hogan, Rosa Beddington, Frank Costantini, and Elizabeth Lacy. The field's technological sophistication has grown exponentially but the manual remains the essential practical and theoretical guide for all students, lab technicians, and investigators who work with mice.
Download and read online PCR primer in PDF and EPUB This second edition of a much-praised and widely used manual has been entirely revised and updated. Each technique is presented with extensive background information, advice, and troubleshooting. All current applications of PCR are covered, in protocols that have the hallmark reliability of the previous edition.
Download and read online BioBuilder in PDF and EPUB Today’s synthetic biologists are in the early stages of engineering living cells to help treat diseases, sense toxic compounds in the environment, and produce valuable drugs. With this manual, you can be part of it. Based on the BioBuilder curriculum, this valuable book provides open-access, modular, hands-on lessons in synthetic biology for secondary and post-secondary classrooms and laboratories. It also serves as an introduction to the field for science and engineering enthusiasts. Developed at MIT in collaboration with award-winning high school teachers, BioBuilder teaches the foundational ideas of the emerging synthetic biology field, as well as key aspects of biological engineering that researchers are exploring in labs throughout the world. These lessons will empower teachers and students to explore and be part of solving persistent real-world challenges. Learn the fundamentals of biodesign and DNA engineering Explore important ethical issues raised by examples of synthetic biology Investigate the BioBuilder labs that probe the design-build-test cycle Test synthetic living systems designed and built by engineers Measure several variants of an enzyme-generating genetic circuit Model "bacterial photography" that changes a strain’s light sensitivity Build living systems to produce purple or green pigment Optimize baker’s yeast to produce ?-carotene
Download and read online Gene Cloning and DNA Analysis in PDF and EPUB Known world-wide as the standard introductory text to this important and exciting area, the seventh edition of Gene Cloning and DNA Analysis addresses new and growing areas of research whilst retaining the philosophy of the previous editions. Assuming the reader has little prior knowledge of the subject, its importance, the principles of the techniques used and their applications are all carefully laid out, with over 250 clearly presented four-colour illustrations. In addition to a number of informative changes to the text throughout the book, the chapters on DNA sequencing and genome studies have been rewritten to reflect the continuing rapid developments in this area of DNA analysis: In depth description of the next generation sequencing methods and descriptions of their applications in studying genomes and transcriptomes New material on the use of ChiP-seq to locate protein-binding sites Extended coverage of the strategies used to assemble genome sequences Description of how the Neanderthal genome has been sequenced and what that sequence tells us about interbreeding between Neanderthals and Homo sapiens Gene Cloning and DNA Analysis remains an essential introductory text to a wide range of biological sciences students; including genetics and genomics, molecular biology, biochemistry, immunology and applied biology. It is also a perfect introductory text for any professional needing to learn the basics of the subject. All libraries in universities where medical, life and biological sciences are studied and taught should have copies available on their shelves.