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 Synthetic Biology in PDF and EPUB Synthetic Biology: A Lab Manual is the first manual for laboratory work in the new and rapidly expanding field of synthetic biology. Aimed at non-specialists, it details protocols central to synthetic biology in both education and research. In addition, it provides all the information that teachers and students from high schools and tertiary institutions need for a colorful lab course in bacterial synthetic biology using chromoproteins and designer antisense RNAs. As a bonus, practical material is provided for students of the annual international Genetically Engineered Machine (iGEM) competition. The manual is based upon a highly successful course at Sweden's Uppsala University and is coauthored by one of the pioneers of synthetic biology and two bioengineering postgraduate students. An inspiring foreword is written by another pioneer in the field, Harvard's George Church: “Synthetic biology is to early recombinant DNA as a genome is to a gene. Is there anything that SynBio will not impact? There was no doubt that the field of SynBio needed ‘A Lab Manual’ such as the one that you now hold in your hands.” Read about the interview with the authors! Contents:Introduction:What is Synthetic Biology, Exactly?The iGEM OutbreakA Synthetic Biology Lab ManualGenes, Chromoproteins and Antisense RNAs:E. coli DNA: Chromosomes, Plasmids and Copy NumberCoupling of Transcription and Translation in BacteriaPromoter and Terminator for TranscriptionRibosome Binding Site (RBS)Codon BiasChromoproteinsSmall Regulatory RNAs (sRNAs)Lab Rooms and Equipment:The Physical Lab SpacesEquipmentSafety is Priority #1:FiresChemicalsBiological Safety and DisposalDangerous EquipmentLab Course Projects:Time and ResourcesProject Overview and Learning ObjectivesThe Lab NotebookLab Section 1. Preparation of Chemical Solutions and Agar PlatesLab Section 2. Coloring Bacteria by Adding a Promoter to a Chromoprotein GeneLab Section 3: Rational Engineering of Chromoprotein Expression LevelLab Section 4. Other ExperimentsThe “Dreaded” ExamProtocols:IntroductionProtocol 1. Preparation of Solutions and Agar PlatesProtocol 2. Overnight Cultures with Antibiotics, and Glycerol StocksProtocol 3. BioBrick™ 3A Assembly and Gel AnalysisProtocol 4. Agarose Gel ElectrophoresisProtocol 5. Preparation of Competent E. coli Cells Using CaCl2Protocol 6. Transformation of CaCl2-Competent E. coli CellsProtocol 7. Bacterial Re-Streaking TechniquesProtocol 8. Lysis of E. coli Cells with LysozymeProtocol 9. Polymerase Chain Reaction (PCR)Protocol 10. Inverse PCR MutagenesisProtocol 11. Colony PCRProtocol 12. Gibson AssemblyAdvanced Methods:Flow Cytometry and Cell SortingRecombination in Plasmids and the ChromosomeElectrocompetent CellsThe International Genetically Engineered Machine (iGEM) Competition:How to Start an iGEM TeamUppsala iGEM 2011 — Show Color with ColorUppsala iGEM 2012 — Resistance is FutileUppsala iGEM 2013 — Lactonutritious — It's DeliciousAppendices Readership: Students and researchers in biotechnology, cell/molecular biology and genetics. Keywords:Synthetic Biology;Genetic Engineering;DNA Cloning;Polymerase Chain Reaction;Molecular Biology;Laboratory;Biobrick;Chromoprotein;Manual;CourseReviews: It provides an introductory workflow for common techniques and would be of definite use to educators wanting to bring synthetic biology into the classroom or for motivated undergraduate students with the desire to start thir own iGEM team.” Synthetic Biology See Full Review
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 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 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 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 Proteins and Proteomics in PDF and EPUB Describes the principles, analytical methods, and protocols involved in the study of proteomics, covering the separation of proteins, liquid chromatography, and sequence analysis.
Download and read online Modern Prometheus in PDF and EPUB This book tells the dramatic story of Crispr and the potential impact of this gene-editing technology.
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.
Download and read online PCR Primer A Laboratory Manual Second Edition 2003 Paperback in PDF and EPUB This second edition of a practical manual has been entirely revised and updated. Each technique is presented with extensive background information, advice and troubleshooting. All contemporary applications of PCR are covered, in protocols that have the hallmark reliability of the previous edition.
Download and read online Manipulating the Mouse Embryo in PDF and EPUB Provides information and guidelines for developing a mouse colony and conducting experiments, including proper protocols, step-by-step procedures, and analysis strategies.
Download and read online Genetics of Complex Human Diseases in PDF and EPUB A handbook explaining suitable study designs, measurement of risk factors, and data reduction in the genetic analysis of human disorders.