Download or read book RT PCR Protocols written by Nicola King and published by Springer Science & Business Media. This book was released on 2008-02-04 with total page 370 pages. Available in PDF, EPUB and Kindle. Book excerpt: Until the mid 1980s, the detection and quantification of a specific mRNA was a difficult task, usually only undertaken by a skilled molecular biologist. With the advent of PCR, it became possible to amplify specific mRNA, after first converting the mRNA to cDNA via reverse transcriptase. The arrival of this technique—termed reverse transcription-PCR (RT-PCR)—meant that mRNA suddenly became amenable to rapid and sensitive analysis, without the need for advanced training in molecular biology. This new accessibility of mRNA, which has been facilitated by the rapid accumulation of sequence data for human mRNAs, means that every biomedical researcher can now include measurement of specific mRNA expression as a routine component of his/her research plans. In view of the ubiquity of the use of standard RT-PCR, the main objective of RT-PCR Protocols is essentially to provide novel, useful applications of RT-PCR. These include some useful adaptations and applications that could be relevant to the wider research community who are already familiar with the basic RT-PCR protocol. For example, a variety of different adaptations are described that have been employed to obtain quantitative data from RT-PCR. Quantitative RT-PCR provides the ability to accurately measure changes/imb- ances in specific mRNA expression between normal and diseased tissues.
Download or read book RT PCR Protocols written by Nicola King and published by Humana Press. This book was released on 2010-04-06 with total page 341 pages. Available in PDF, EPUB and Kindle. Book excerpt: Once a tedious, highly skilled operation, reverse-transcription polymerase chain reaction (RT-PCR) has become a routine and invaluable technique used in most laboratories. In RT-PCR Protocols, Second Edition, expert researchers fully update the technologies presented in the popular previous edition, such as competitive RT-PCR, nested RT-PCR, RT-PCR from single cells, and RT-PCR for cloning. In addition, newer technologies are also explored, including multiplex RT-PCR, RT-LATE-PCR, and the greatly advanced field of real-time quantitative RT-PCR, while recent advances in creating the optimum RT-PCR reaction, e.g. RNA extraction, primer design, and reverse transcription, end the book with their indispensable input. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include brief introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes sections, highlighting tips on troubleshooting and avoiding known pitfalls. User friendly and up-to-date, RT-PCR Protocols, Second Edition acts as a handy companion to scientists from numerous diverse backgrounds who wish to explore further the marvels of gene expression.
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 Quantitative Real Time PCR written by Roberto Biassoni and published by Humana. This book was released on 2014-04-17 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Quantitative Real-Time PCR: Methods and Protocols focuses on different applications of qPCR ranging from microbiological detections (both viral and bacterial) to pathological applications. Several chapters deal with quality issues which regard the quality of starting material, the knowledge of the minimal information required to both perform an assay and to set the experimental plan, while the others focus on translational medicine applications that are ordered following an approximate logical order of their medical application. The last part of the book gives you an idea of an emerging digital PCR technique that is a unique qPCR approach for measuring nucleic acid, particularly suited for low level detection and to develop non-invasive diagnosis. Written for the Methods in Molecular Biology series, most chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, laboratory protocols and tips on troubleshooting and avoiding known pitfalls. Practical and authoritative, Quantitative Real-Time PCR: Methods and Protocols aims to aid researchers seeking to devise new qPCR-based approaches related to his or her area of investigation.
Download or read book RT PCR Protocols written by Nicola King and published by Humana. This book was released on 2016-08-23 with total page 341 pages. Available in PDF, EPUB and Kindle. Book excerpt: Once a tedious, highly skilled operation, reverse-transcription polymerase chain reaction (RT-PCR) has become a routine and invaluable technique used in most laboratories. In RT-PCR Protocols, Second Edition, expert researchers fully update the technologies presented in the popular previous edition, such as competitive RT-PCR, nested RT-PCR, RT-PCR from single cells, and RT-PCR for cloning. In addition, newer technologies are also explored, including multiplex RT-PCR, RT-LATE-PCR, and the greatly advanced field of real-time quantitative RT-PCR, while recent advances in creating the optimum RT-PCR reaction, e.g. RNA extraction, primer design, and reverse transcription, end the book with their indispensable input. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include brief introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes sections, highlighting tips on troubleshooting and avoiding known pitfalls. User friendly and up-to-date, RT-PCR Protocols, Second Edition acts as a handy companion to scientists from numerous diverse backgrounds who wish to explore further the marvels of gene expression.
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 Learning from SARS written by Institute of Medicine and published by National Academies Press. This book was released on 2004-04-26 with total page 376 pages. Available in PDF, EPUB and Kindle. Book excerpt: The emergence of severe acute respiratory syndrome (SARS) in late 2002 and 2003 challenged the global public health community to confront a novel epidemic that spread rapidly from its origins in southern China until it had reached more than 25 other countries within a matter of months. In addition to the number of patients infected with the SARS virus, the disease had profound economic and political repercussions in many of the affected regions. Recent reports of isolated new SARS cases and a fear that the disease could reemerge and spread have put public health officials on high alert for any indications of possible new outbreaks. This report examines the response to SARS by public health systems in individual countries, the biology of the SARS coronavirus and related coronaviruses in animals, the economic and political fallout of the SARS epidemic, quarantine law and other public health measures that apply to combating infectious diseases, and the role of international organizations and scientific cooperation in halting the spread of SARS. The report provides an illuminating survey of findings from the epidemic, along with an assessment of what might be needed in order to contain any future outbreaks of SARS or other emerging infections.
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 Real Time PCR written by Kirstin J. Edwards and published by Taylor & Francis. This book was released on 2004 with total page 362 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Real time PCR written by M Dorak and published by Garland Science. This book was released on 2007-01-24 with total page 484 pages. Available in PDF, EPUB and Kindle. Book excerpt: With a variety of detection chemistries, an increasing number of platforms, multiple choices for analytical methods and the jargon emerging along with these developments, real-time PCR is facing the risk of becoming an intimidating method, especially for beginners. Real-time PCR provides the basics, explains how they are exploited to run a real-time PCR assay, how the assays are run and where these assays are informative in real life. It addresses the most practical aspects of the techniques with the emphasis on 'how to do it in the laboratory'. Keeping with the spirit of the Advanced Methods Series, most chapters provide an experimental protocol as an example of a specific assay.
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 Differential Display Reverse Transcription PCR DDRT PCR written by Sergio Colonna-Romano and published by Springer Science & Business Media. This book was released on 2012-12-06 with total page 135 pages. Available in PDF, EPUB and Kindle. Book excerpt: Identification of differentially expressed genes is one of the major challenges in molecular biology. Several techniques allow the cloning of such sequences. However, methods such as RNA subtraction or differential hybridization are time-consuming and require large amounts of mRNA. Recently, a new approach has successfully been developed: Differential-Display Reverse Transcription-PCR (DDRT-PCR). This technique has been proven to be highly effective in identifying sequences that are differentially expressed in various cell types. The most striking advantage is, however, that only nanograms of total RNA are sufficient. Thus every mRNA species expressed in the cell system can be investigated, even those present at very low levels.
Download or read book PCR Protocols written by Bruce A. White and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 397 pages. Available in PDF, EPUB and Kindle. Book excerpt: PCR has been successfully utilized in every facet of basic, cli- cal, and applied studies of the life sciences, and the impact that PCR has had on life science research is already staggering. C- comitant with the essentially universal use of PCR has been the creative and explosive development of a wide range of PCR-based techniques and applications. These increasingly numerous pro- cols have each had the general effect of facilitating and acceler- ing research. Because PCR technology is relatively easy and inexpensive, PCR applications are well within the reach of every research lab. In this sense, PCR has become the "equalizer" between "small" and "big" labs, since its use makes certain projects, especially those related to molecular cloning, now far more feasible for the small lab with a modest budget. This new volume on PCR Protocols does not attempt the impossible task of representing all PCR-based protocols. Rather, it presents a range of protocols, both analytical and preparative, that provide a solid base of knowledge on the use of PCR in many c- mon research problems. The first six chapters provide some basic information on how to get started. Chapters 7-19 represent primarily analytical uses of PCR, both for simple DNA and RNA detection, as well as for more complex analyses of nucleic acid (e. g. , DNA footprin ting, RNA splice site localization). The remaining chapters represent "synthetic," or preparative, uses of PCR.
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 Rapid Cycle Real Time PCR Methods and Applications written by W. Dietmaier and published by Springer Science & Business Media. This book was released on 2013-06-29 with total page 200 pages. Available in PDF, EPUB and Kindle. Book excerpt: Rapid-Cycle Real-Time PCR is a powerful technique for nucleic acid amplification and analysis that often requires less than half an hour to perform. Samples are amplified by rapid-cycle PCR followed by immediate melting curve analysis in the same instrument. Melting curve analysis of PCR products with SYBR Green I often allows product identification without gel electrophoresis. Furthermore, in the presence of fluorescent hybridization probes, melting curves provide "dynamic dot blots" for fine sequence analysis, including single nucleotide polymorphisms (SNPs). The method is often cited as the most versatile, efficient method for nucleic acid analysis in research and diagnostics in the fields of genetics and oncology. Molecular diagnostics has never been easier!
Download or read book Avian Influenza Virus written by Erica Spackman and published by Springer Science & Business Media. This book was released on 2008-02-28 with total page 147 pages. Available in PDF, EPUB and Kindle. Book excerpt: With the growing global fear of a major pandemic, avian influenza (AI) virus research has greatly increased in importance. In Avian Influenza Virus, an expert team of researchers and diagnosticians examine the fundamental, yet essential, virological methods for AI virus research and diagnostics as well as some of the newest molecular procedures currently used for basic and applied research. They present exciting, cutting-edge new methods that focus both on studying the virus itself and on work with avian hosts, an area greatly lacking in research.
Download or read book PCR Protocols in Molecular Toxicology written by John P. Vanden Heuvel and published by CRC Press. This book was released on 2019-05-07 with total page 271 pages. Available in PDF, EPUB and Kindle. Book excerpt: Molecular toxicology is an emerging discipline that utilizes molecular and cell biology to understand how drugs and chemicals result in their unwanted effects. PCR Protocols in Molecular Toxicology is a practical guide to the use of polymerase chain reaction (PCR) to help examine, on a molecular and cellular level, how toxic responses are manifested. It offers a basic understanding of PCR and its optimization, as well as describing specific, high-impact areas of molecular toxicology and recent advances. The following techniques are described in detail: Quantitative reverse transcriptase PCR and methods to examine gene expression Differential display cloning Cloning and library screening by PCR Genotype and polymorphism analysis of drug and toxicant metabolizing enzymes Basic, non-PCR based molecular biology methods PCR Protocols in Molecular Toxicology will aid both novices and experienced PCR practitioners in using PCR to its fullest potential.
