Download or read book PCR Troubleshooting and Optimization written by Suzanne Kennedy and published by . This book was released on 2011 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: This book discusses the strategies for preparing effective controls and standards for PCR, when they should be employed and how to interpret the information they provide. It highlights the significance of optimization for efficiency, precision and sensitivity of PCR methodology and provides essential guidance on how to troubleshoot inefficient reactions. Experts in PCR describe design and optimization techniques, discuss the use of appropriate controls, explain the significance of standard curves and explore the principles and strategies required for effective troubleshooting. Authors highligh.
Download or read book PCR Troubleshooting and Optimization written by Suzanne Kennedy and published by Caister Academic Press Limited. This book was released on 2011 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: The polymerase chain reaction (PCR) is a fundamental tool in scientific research and clinical testing. Real-time PCR, combining both amplification and detection in one instrument, is a rapid and accurate method for nucleic acid detection and quantification. Although PCR is a very powerful technique, the results achieved are valid only if the appropriate controls have been employed. In addition, proper optimization of PCR conditions is required for the generation of specific, repeatable, reproducible, and sensitive data. This book discusses the strategies for preparing effective controls and standards for PCR, when they should be employed, and how to interpret the information they provide. It highlights the significance of optimization for efficiency, precision, and sensitivity of PCR methodology and provides essential guidance on how to troubleshoot inefficient reactions. Experts in PCR describe design and optimization techniques, discuss the use of appropriate controls, explain the significance of standard curves, and explore the principles and strategies required for effective troubleshooting. The book highlights the importance of sample preparation and quality, primer design, controlling inhibitors, avoiding amplicon and environmental contamination, optimizing reagent quality and concentration, and modifying the thermal cycling protocol for optimal sensitivity and specificity. In addition, specific chapters discuss the history of PCR, the choice of instrumentation, the applications of PCR in metagenomics, high resolution melting analysis, the MIQE guidelines, and PCR at the microliter scale. The strategies, tips and advice contained in this concise volume will enable the scientist to optimize and effectively troubleshoot a wide range of techniques, including PCR, reverse transcriptase PCR, real-time PCR, and quantitative PCR. It will be an essential book for anyone using PCR technology.
Download or read book PCR Protocols written by Michael A. Innis and published by Academic Press. This book was released on 2012-12-02 with total page 501 pages. Available in PDF, EPUB and Kindle. Book excerpt: The correct procedures you need for frustration-free PCR methods and applications are contained in this complete, step-by-step, clearly written, inexpensive manual. Avoid contamination--with specific instructions on setting up your lab Avoid cumbersome molecular biological techniques Discover new applications
Download or read book PCR Guru written by Ayaz Najafov and published by Academic Press. This book was released on 2016-11-28 with total page 108 pages. Available in PDF, EPUB and Kindle. Book excerpt: PCR Guru: An Ultimate Benchtop Reference for Molecular Biologists is provides researchers in molecular biology with a handy reference for approaching and solving challenging problems associated with PCR setup and optimization. As a laboratory guide, it emphasizes the technical aspects of employing PCR as a tool in molecular biology laboratories. The book covers the history of PCR and the basic science underlying it. It then discusses PCR at the bench level, starting with detailed description and tips on primer design, and continuing with the standard protocols used to perform PCR. Provides troubleshooting tips for various types of modifications of standard protocols Contains unique “Good Practices and Tips that are indispensable for the beginner and expert alike Features “Special Cases with applications of PCR, optimization, and troubleshooting Includes detailed appendices with tables, figures, and key protocols Organized as a systematic, concentrated resource to save time when addressing a PCR problem
Download or read book PCR Troubleshooting written by Michael L. Altshuler and published by Horizon Scientific Press. This book was released on 2006 with total page 80 pages. Available in PDF, EPUB and Kindle. Book excerpt: This unique polymerase chain reaction (PCR) troubleshooting guide is an essential companion for readers with some experience in PCR. The book discusses the many and varied problems encountered with PCR, together with tips, advice, and procedures to obviate rather than overcome the PCR problems. The advice in PCR Troubleshooting is invaluable.
Download or read book Gene Quantification written by Francois Ferre and published by Springer Science & Business Media. This book was released on 2012-12-06 with total page 379 pages. Available in PDF, EPUB and Kindle. Book excerpt: Geneticists and molecular biologists have been interested in quantifying genes and their products for many years and for various reasons (Bishop, 1974). Early molecular methods were based on molecular hybridization, and were devised shortly after Marmur and Doty (1961) first showed that denaturation of the double helix could be reversed - that the process of molecular reassociation was exquisitely sequence dependent. Gillespie and Spiegelman (1965) developed a way of using the method to titrate the number of copies of a probe within a target sequence in which the target sequence was fixed to a membrane support prior to hybridization with the probe - typically a RNA. Thus, this was a precursor to many of the methods still in use, and indeed under development, today. Early examples of the application of these methods included the measurement of the copy numbers in gene families such as the ribosomal genes and the immunoglo bulin family. Amplification of genes in tumors and in response to drug treatment was discovered by this method. In the same period, methods were invented for estimating gene num bers based on the kinetics of the reassociation process - the so-called Cot analysis. This method, which exploits the dependence of the rate of reassociation on the concentration of the two strands, revealed the presence of repeated sequences in the DNA of higher eukaryotes (Britten and Kohne, 1968). An adaptation to RNA, Rot analysis (Melli and Bishop, 1969), was used to measure the abundance of RNAs in a mixed population.
Download or read book Molecular Diagnostic PCR Handbook written by Gerrit J. Viljoen and published by Springer Science & Business Media. This book was released on 2005-12-06 with total page 325 pages. Available in PDF, EPUB and Kindle. Book excerpt: PREFACE The Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture is involved in agricultural research and development and assists Member States of FAO and IAEA in improving strategies to ensure food security through the use of nuclear techniques and related biotechnologies, where such techniques have a valuable and often unique role. In particular, molecular diagnostic methods have rapidly evolved in the past twenty years, since the advent of the Polymerase Chain Reaction (PCR). They are used in a wide range of agricultural areas such as, improving soil and water management; producing better crop varieties; diagnosing plant and animal diseases; controlling insect pests and improving food quality and safety. The uses of nucleic acid-directed methods have increased significantly in the past five years and have made important contributions to disease control country programmes for improving national and international trade. These developments include the more routine use of PCR as a diagnostic tool in veterinary diagnostic laboratories. However, there are many problems associated with the transfer and particularly, the application of this technology. These include lack of consideration of: the establishment of quality-assured procedures, the required set-up of the laboratory and the proper training of staff. This can lead to a situation where results are not assured. This book gives a comprehensive account of the practical aspects of PCR and strong consideration is given to ensure its optimal use in a laboratory environment. This includes the setting-up of a PCR laboratory; Good Laboratory Practice and standardised of PCR protocols.
Download or read book PCR Primer written by Carl W. Dieffenbach and published by CSHL Press. This book was released on 2003 with total page 544 pages. Available in PDF, EPUB and Kindle. Book excerpt: The Polymerase Chain Reaction (PCR) technique was invented nearly 20 years ago. Its subsequent variations and applications were many and varied, and today molecular biology, clinical, and forensic laboratories make almost daily use of PCR. This second edition of the much-praised PCR Primer: A Laboratory Manual updates the tried-and-true methods and presents the advances made in the 10 years since the first edition. After introducing the basics for PCR and methods of sample preparation, PCR Primer provides laboratory-tested protocols for RT-PCR methods, detection of PCR products, analysis of differential expression, cloning, and mutagenesis. These step-by-step methods include extensive background information, as well as valuable troubleshooting information provided by the leading experts in this technology. This manual is a comprehensive and reliable source of the full range of PCR methods for novices and experienced investigators alike.
Download or read book PCR Technology written by Henry Erlich and published by Springer. This book was released on 2015-12-31 with total page 246 pages. Available in PDF, EPUB and Kindle. Book excerpt: This is an introduction to the methods and applications of polymerase chain reaction (PCR) technology, a technology developed by Erlich's group at Cetus and Cetus, and is expected to be used in all biology laboratories worldwide within the next few years.
Download or read book Current Protocols Essential Laboratory Techniques written by Sean R. Gallagher and published by John Wiley & Sons. This book was released on 2012-03-19 with total page 679 pages. Available in PDF, EPUB and Kindle. Book excerpt: The latest title from the acclaimed Current Protocols series, Current Protocols Essential Laboratory Techniques, 2e provides the new researcher with the skills and understanding of the fundamental laboratory procedures necessary to run successful experiments, solve problems, and become a productive member of the modern life science laboratory. From covering the basic skills such as measurement, preparation of reagents and use of basic instrumentation to the more advanced techniques such as blotting, chromatography and real-time PCR, this book will serve as a practical reference manual for any life science researcher. Written by a combination of distinguished investigators and outstanding faculty, Current Protocols Essential Laboratory Techniques, 2e is the cornerstone on which the beginning scientist can develop the skills for a successful research career.
Download or read book PCR Protocols written by John M. S. Bartlett and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 1083 pages. Available in PDF, EPUB and Kindle. Book excerpt: In this new edition, the editors have thoroughly updated and dramatically expanded the number of protocols to take advantage of the newest technologies used in all branches of research and clinical medicine today. These proven methods include real time PCR, SNP analysis, nested PCR, direct PCR, and long range PCR. Among the highlights are chapters on genome profiling by SAGE, differential display and chip technologies, the amplification of whole genome DNA by random degenerate oligonucleotide PCR, and the refinement of PCR methods for the analysis of fragmented DNA from fixed tissues. Each fully tested protocol is described in step-by-step detail by an established expert in the field and includes a background introduction outlining the principle behind the technique, equipment and reagent lists, tips on trouble shooting and avoiding known pitfalls, and, where needed, a discussion of the interpretation and use of results.
Download or read book PCR Primer Design written by Chhandak Basu and published by Humana. This book was released on 2016-10-08 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume provides an overview on design PCR primers for successful DNA amplification. Chapters focus on primer design strategies for quantitative PCR, in silico PCR primer design, and primer design using software. 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 easily accessible, PCR Primer Design, Second Edition seeks to aid molecular biology students, researchers, professors and PCR enthusiasts.
Download or read book PCR Technology written by Tania Nolan and published by CRC Press. This book was released on 2013-06-13 with total page 461 pages. Available in PDF, EPUB and Kindle. Book excerpt: PCR’s simplicity as a molecular technique is, in some ways, responsible for the huge amount of innovation that surrounds it, as researchers continually think of new ways to tweak, adapt, and re-formulate concepts and applications. PCR Technology: Current Innovations, Third Edition is a collection of novel methods, insights, and points of view that provides a critical and timely reference point for anyone wishing to use this technology. Topics in this forward-thinking volume include: The purification and handling of PCR templates The effect of the manufacture and purification of the oligonucleotide on PCR behavior Optimum buffer composition Probe options The design and optimization of qPCR assays Issues surrounding the development and refinement of instrumentation Effective controls to protect against uncertainties due to reaction variability Covering all aspects of PCR and real-time PCR, the book contains detailed protocols that make it suitable as both a reference and an instruction manual. Each chapter presents detailed guidelines as well as helpful hints and tips supplied by authors who are recognized experts in their fields. In addition to descriptions of current technology and best practices, the book also provides information about new developments in the PCR arena.
Download or read book PCR written by Lucília Domingues and published by Springer Nature. This book was released on 2023-09-23 with total page 255 pages. Available in PDF, EPUB and Kindle. Book excerpt: This second volume focuses on PCR methods and PCR application specificities to the biotechnology and bioengineering field. New and updated chapters detail real-time PCR protocols, synthetic biology applications, pathogen detection, microfluidics, digital, multiplex detection recent advances. 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 key tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, PCR: Methods and Protocols, Second Edition aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge.
Download or read book The Polymerase Chain Reaction written by Kary B. Mullis and published by Springer Science & Business Media. This book was released on 2012-02-02 with total page 464 pages. Available in PDF, EPUB and Kindle. Book excerpt: James D. Watson When, in late March of 1953, Francis Crick and I came to write the first Nature paper describing the double helical structure of the DNA molecule, Francis had wanted to include a lengthy discussion of the genetic implications of a molecule whose struc ture we had divined from a minimum of experimental data and on theoretical argu ments based on physical principles. But I felt that this might be tempting fate, given that we had not yet seen the detailed evidence from King's College. Nevertheless, we reached a compromise and decided to include a sentence that pointed to the biological significance of the molecule's key feature-the complementary pairing of the bases. "It has not escaped our notice," Francis wrote, "that the specific pairing that we have postulated immediately suggests a possible copying mechanism for the genetic material." By May, when we were writing the second Nature paper, I was more confident that the proposed structure was at the very least substantially correct, so that this second paper contains a discussion of molecular self-duplication using templates or molds. We pointed out that, as a consequence of base pairing, a DNA molecule has two chains that are complementary to each other. Each chain could then act ". . . as a template for the formation on itself of a new companion chain, so that eventually we shall have two pairs of chains, where we only had one before" and, moreover, " ...
Download or read book PCR Cloning Protocols written by Bruce Alan White and published by . This book was released on 1997 with total page 490 pages. Available in PDF, EPUB and Kindle. Book excerpt: Distinguished scientists and researchers present a comprehensive collection of current preparative PCR techniques that can be used in cloning and modifying DNA and cDNA. Topics include performing and optimizing PCR (including long PCR), cloning PCR products, cloning unknown neighboring DNA, and library construction and screening. Also covered are mutagenesis, recombination, and in vitro selection, differential and subtractive approaches to cDNA analysis and screening, and cloning members of gene families. The techniques bring to both new and established researchers the power to apply PCR-based methodology to the cloning and modification of DNA, either through innovative protocols or by fostering individual creativity to modify and customize the protocols to best fit their own needs.
Download or read book Plant Functional Genomics written by Erich Grotewold and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 443 pages. Available in PDF, EPUB and Kindle. Book excerpt: Functional genomics is a young discipline whose origin can be traced back to the late 1980s and early 1990s, when molecular tools became available to determine the cellular functions of genes. Today, functional genomics is p- ceived as the analysis, often large-scale, that bridges the structure and organi- tion of genomes and the assessment of gene function. The completion in 2000 of the genome sequence of Arabidopsis thaliana has created a number of new and exciting challenges in plant functional genomics. The immediate task for the plant biology community is to establish the functions of the approximately 25,000 genes present in this model plant. One major issue that will remain even after this formidable task is c- pleted is establishing to what degree our understanding of the genome of one model organism, such as the dicot Arabidopsis, provides insight into the or- nization and function of genes in other plants. The genome sequence of rice, completed in 2002 as a result of the synergistic interaction of the private and public sectors, promises to significantly enrich our knowledge of the general organization of plant genomes. However, the tools available to investigate gene function in rice are lagging behind those offered by other model plant systems. Approaches available to investigate gene function become even more limited for plants other than the model systems of Arabidopsis, rice, and maize.
