Download or read book PCR 3 written by C. Simon Herrington and published by Oxford University Press. This book was released on 1997-10-30 with total page 223 pages. Available in PDF, EPUB and Kindle. Book excerpt: PCR in situ hybridization allows the detection of specific nucleic acid sequences and their distribution in the cell. It combines two powerful techniquesin situ hybridization (ISH), which allows cellular localization of DNA and RNA sequences in cells and tissues, and the polymerase chain reaction (PCR), which allows reproducible amplification of rare nucleic acid sequences. The combined technique and its variants greatly enhance the sensitivity of in situ hybridization and add morphological localization to the sensitivity of PCR. Such techniques have enormous potential for research and diagnosis but problems with reproducibility and reliability are often encountered. This book overcomes these problems by describing the key procedures in step-by-step detail and by providing the essential advice needed for success. Topics include: DNA in situ PCR and DNA PCR in situ hybridization (PCR ISH) for the detection of DNA targets in cells; reverse trancriptase in situ PCR (RT-PCR) and RT-PCR ISH for the detection of RNA targets; and PRINS (primed in situ synthesis) for chromosomal analysis in interphase nuclei and metaphase chromosome spreads. There are further chapters on fixation of tissues for PCR, selective ultraviolet radiation fractionation (SURF), application of in situ PCR to human tissues, applications and modifications of PCR-ISH, and automation of in situ amplification. PCR In Situ Hybridization is a unique and timely collection of well-tested protocols for the amplification of DNA and RNA in cells and tissues, drawing on the accumulated knowledge and experience of leading exponents of these techniques. For each topic covered, the authors provide detailed guidance on the key steps in the protocols, numerous hints and tips for success, and advice on trouble-shooting. PCR In Situ Hybridization will be invaluable to molecular biologists, pathologists, geneticists, and all those seeking to perform in situ analyses of nucleic acid molecules.
Download or read book PCR 3 written by C. S. Herrington and published by . This book was released on 1998 with total page 228 pages. Available in PDF, EPUB and Kindle. Book excerpt: PCR in situ hybridization allows the detection of specific nucleic acid sequences and their distribution in the cell. It combines two powerful techniquesin situ hybridization (ISH), which allows cellular localization of DNA and RNA sequences in cells and tissues, and the polymerase chainreaction (PCR), which allows reproducible amplification of rare nucleic acid sequences. The combined technique and its variants greatly enhance the sensitivity of in situ hybridization and add morphological localization to the sensitivity of PCR. Such techniques have enormous potential for researchand diagnosis but problems with reproducibility and reliability are often encountered. This book overcomes these problems by describing the key procedures in step-by-step detail and by providing the essential advice needed for success. Topics include: DNA in situ PCR and DNA PCR in situhybridization (PCR ISH) for the detection of DNA targets in cells; reverse trancriptase in situ PCR (RT-PCR) and RT-PCR ISH for the detection of RNA targets; and PRINS (primed in situ synthesis) for chromosomal analysis in interphase nuclei and metaphase chromosome spreads. There are furtherchapters on fixation of tissues for PCR, selective ultraviolet radiation fractionation (SURF), application of in situ PCR to human tissues, applications and modifications of PCR-ISH, and automation of in situ amplification. PCR In Situ Hybridization is a unique and timely collection of well-testedprotocols for the amplification of DNA and RNA in cells and tissues, drawing on the accumulated knowledge and experience of leading exponents of these techniques. For each topic covered, the authors provide detailed guidance on the key steps in the protocols, numerous hints and tips for success,and advice on trouble-shooting. PCR In Situ Hybridization will be invaluable to molecular biologists, pathologists, geneticists, and all those seeking to perform in situ analyses of nucleic acid molecules.
Download or read book PCR RT PCR in situ written by Gerard Morel and published by CRC Press. This book was released on 2002-09-27 with total page 434 pages. Available in PDF, EPUB and Kindle. Book excerpt: Although the polymerase chain reaction has revolutionized genetic analysis by amplifying rare nucleic acid sequences, the in situ application is the only method that allows the localization of amplified signal within tissue structure. The applications of in situ polymerase chain reaction have greatly enhanced the field of investigation in many disc
Download or read book In Situ PCR Techniques written by Omar Bagasra and published by Wiley-Liss. This book was released on 1997-07-04 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book describes comprehensive step-by-step protocols for the delineation of genetic amplification and histological detection techniques. Each procedure has been tested and validated for its sensitivity, precision, and reproducibility, and the authors give advice on the design of primers for PCR applications and on optimizing these protocols for use with plant, insect, and prokaryotic cells.
Download or read book PRINS and In Situ PCR Protocols written by Franck Pellestor and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 257 pages. Available in PDF, EPUB and Kindle. Book excerpt: The in situ hybridization and PCR technologies are now well-established molecular techniques for studying chromosomal aneuploidy and rearran- ments, gene localization and expression, and genomic organization. Over the last decade, we have seen increasing applications in these fields. By combining the high sensitivity of the PCR reaction and the cytological localization of target sequences, both PRINS and in situ PCR techniques have provided highly powerful complements to FISH for in situ cellular and molecular investigations. Both these approaches have several advantages in terms of sensitivity and specificity, owing to the use of primers and to the fast kinetics of annealing and elongation reactions in situ. In the first edition of PRINS and In Situ PCR Protocols edited by John R. Gosden, experts in the field presented in detail a variety of applications of PRINS and in situ PCR techniques, in a wide range of clinical conditions. Since the publication of this successful reference book, there have been s- nificant improvements in in situ detection techniques. This completely revised and updated second edition presents a compreh- sive selection of new procedures developed in the field of PRINS and in situ PCR technologies. The book has two sections. Part I, Basic Methodology, contains chapters that provide useful protocols for many variations of PRINS and in situ PCR, including a new fast multicolor PRINS method, and protocols for PRINS detection of unique sequences in situ.
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 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 Ovarian Cancer written by John M. S. Bartlett and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 787 pages. Available in PDF, EPUB and Kindle. Book excerpt: If there is one aspect of current cancer research that represents a major ch- lenge in both novice and experienced researchers, it is the rapid advance in our understanding of the disease. Researchers can be required to switch from analysis of gene expression to kinetics of protein activation, from genetic studies to the analysis of protein funtion. Cancers are highly complex disease systems and researchers aiming to understand the functioning of cancer systems require access to a wide range of laboratory techiques from a broad range of research disciplines. Increasingly, however, published methods are incomplete or refer back to a series of previous publications each containing only a small part of the complete pro- col. The aim of Ovarian Cancer: Methods and Protocols is to provide for ovarian cancer researchers in the first instance, a laboratory handbook that will facilitate research into cancer systems by providing a series of expert protocols, with proven efficacy, across a broad range of technical expertise. Thus, there are sections on tumor genetics and cellular signal transduction, as well as sections on apoptosis and RNA analysis. The value of Ovarian Cancer: Methods and Protocols to the ovarian cancer researcher will, I trust, be considerably enhanced by (1) the provision of a series of overviews relating to the biology, diagnosis, and treatment of this important neoplasm, and (2) the provision of a series of technical overviews introducing each part that provides an expert review of the applications and pitfalls of the various techniques included.
Download or read book Principles and Technical Aspects of PCR Amplification written by Elizabeth van Pelt-Verkuil and published by Springer Science & Business Media. This book was released on 2008-03-14 with total page 333 pages. Available in PDF, EPUB and Kindle. Book excerpt: Kary Mullis was awarded a Nobel Prize for inventing the PCR technique more than a decade ago in 1993. Since its "discovery", multiple adaptations and variations of the standard PCR technique have been described. This publication aims to provide the reader with a guide to the standard PCR technique and its many available variants, with particular emphasis being placed on the role of these PCR techniques in the clinical diagnostic laboratory (the central theme of this book).
Download or read book In Situ PCR and Related Technology written by Jiang Gu and published by Birkhäuser. This book was released on 2012-02-25 with total page 143 pages. Available in PDF, EPUB and Kindle. Book excerpt: Ever since the introduction of the polymerase chain reaction (peR) in 1986, morphologists, whose interests lie in the analysis of intact tissue structures, have been attempting to adapt this technique to intact cells or tissue sections to detect low copy numbers of DNA or RNA in situ while preserving tissue morphology. The significance of this objective is obvious. A technique finally materialized in 1990 when Dr. Ashley T. Haase and coworkers published results that used multiple prim ers with complementary tails in intact cells. Since then, a number of laboratories have successfully developed their own versions of the technique. In situ peR is now a well-recognized method that permits the detection of minute quantities of DNA or RNA in intact cells or tissue sections. As a result, morphological analysis of those target nucleotide sequences becomes possible. As anticipated, this ad vancement has led to significant improvement in our understanding of many nor mal and abnormal conditions, and its impact is becoming more evident as time passes. In situ peR has the characteristics of a new landmark in morphologic technol ogy-it is scientifically fascinating and technically challenging. In essence, it is a combination of in situ hybridization and conventional peR. The wealth of litera ture, experience and protocols for the two latter techniques can be applied to in situ peR. In situ peR also has its own unique aspects that were not addressed by the other two techniques.
Download or read book PCR Applications written by Michael A. Innis and published by Academic Press. This book was released on 1999-05-11 with total page 589 pages. Available in PDF, EPUB and Kindle. Book excerpt: PCR is the most powerful technique currently used in molecular biology. It enables the scientist to quickly replicate DNA and RNA on the benchtop. From its discovery in the early 80's, PCR has blossomed into a method that enables everything from ready mutation of DNA/RNA to speedy analysis of tens of thousands of nucleotide sequences daily.PCR Applications examines the latest developments in this field. It is the third book in the series, building on the previous publications PCR Protocols and PCR Strategies. The manual discusses techniques that focus on gene discovery, genomics, and DNA array technology, which are contributing factors to the now-occurring bioinformatics boom.Key Features* Focuses on gene discovery, genomics, and DNA array technology* Covers quantitative PCR techniques, including the use of standards and kinetic analysisincludes statistical refinement of primer design parameters* Ilustrates techniques used in microscopic tissue samples, such as single cell PCR, whole cell PCR, laser capture microdissection, and in situ PCREntries provide information on:* Nomenclature* Expression* Sequence analysis* Structure and function* Electrophysiology* Parmacology* Information retrieval
Download or read book PCR in Situ Hybridization written by Gerard J. Nuovo and published by Lippincott Williams & Wilkins. This book was released on 1997 with total page 536 pages. Available in PDF, EPUB and Kindle. Book excerpt: Describes the technique whereby the extreme sensitivity of the polymerase chain reaction (PCR) is combined with the cell localizing ability of in situ hybridization. This revised and updated edition contains chapters on the basics of molecular biology; the nonspecific pathways of PCR; applications of PCR in situ hybridization--human papillomavirus, and HIV-1; and instrumentation. There is also an appendix on reagents for molecular biological analyses. Annotation copyright by Book News, Inc., Portland, OR
Download or read book PCR in Situ Hybridization written by and published by . This book was released on with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Modern Applications of DNA Amplification Techniques written by Dirk Lassner and published by Springer. This book was released on 2013-11-11 with total page 143 pages. Available in PDF, EPUB and Kindle. Book excerpt: In the ten years since the first publication on PCR (Saiki et al. , 1985), this in vitro method of nucleic acid replication and modification has grown to rival in popularity traditional microbiological, genetical und technical procedures for cloning, sequencing, gene detecting and related procedures. To date the PCR literature has emphasized six main areas of application: genetic mapping, detection of mutations, genetic polymorphism, transcriptional splicing and regulation, molecular virology and quantitative procedures. The overwhelming focus of quantification of DNA or RNA by PCR has been on human microbiology and oncological problems. The exquisite sensitivity of PCR gives this method the ability to detect extremely rare DNAs, mRNAs, mRNAs in small numbers of cells or in small amounts of tissue, and mRNAs expressed in mixed-cell populations. However, the exact and accurate quantification of specific nucleic acids in biological samples is in spite of numerous publications in that field still a general problem: during the peR process, an unknown initial number of target sequences are used as a template from which a large quantity of specific product can be obtained. Although the amount of product formed is easy to determine, it is difficult to deduce the initial copy number of the target molecule because the efficiency of the peR is largely unknown.
Download or read book PRINS and In Situ PCR Protocols Methods in Molecular Biology written by Franck Pellestor and published by . This book was released on 2006 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: This completely revised and updated edition takes advantage of the many new developments and applications that have occurred in PRINS and in situ PCR technology, including a new fast multicolor PRINS method for identifying human chromosomes, PRINS protocols for in situ detection of unique sequences and point mutation analysis, a new dideoxy PRINS procedure, and in situ PCR and PRINS protocols for plants. The authors describe in detail various applications of PRINS in human (detection of gene deletions in cancer detection of fetal cells in maternal blood, and assessment of aneuploidy in brain tissues and embryos) as well as plant cells. Readily reproducible in situ PCR techniques are also presented for the detection of cytomegalovirus, and for use in combination with microdissection. In situ RT-PCR techniques are also given for use in plant or cancer 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 Molecular Methods for Virus Detection written by Danny L. Wiedbrauk and published by Elsevier. This book was released on 1995-02-08 with total page 405 pages. Available in PDF, EPUB and Kindle. Book excerpt: Molecular diagnostic procedures have been described in a number of recent books and articles. However, these publications have not focused on virus detection, nor have they provided practical protocols for the newer molecular methods. Written by the inventors or principal developers of these technologies, Molecular Methods for Virus Detection provides both reviews of individual methods and instructions for detecting virus nucleic acid sequences in clinical specimens. Each procedure includes quality assurance protocols that are often ignored by other methodology books. Molecular Methods for Virus Detection provides clinically relevant procedures for many of the newer diagnostic methodologies. - Provides state-of-the-art PCR methods for amplification, quantitation, in situ hybridization, and multiplex reactions - Goes beyond PCR with protocols for 3SR, NASBA, LCR, SDA, and LAT - Covers important virus detection methods such as in situ hybridization; Southern, dot, and slot blots; branched chain signal amplification; and chemiluminescence - Includes quality control information crucial in research and clinical laboratories - Most chapters are written by the inventors and principal developers of the methodologies - Includes color plates, 77 figures, and 18 tables
