Pcr Rt Pcr In Situ
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| Author | : C. Simon Herrington |
| Publisher | : Oxford University Press |
| Total Pages | : 223 |
| Release | : 1997-10-30 |
| Genre | : Science |
| ISBN | : 0191565598 |
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.
| Author | : Gerard Morel |
| Publisher | : CRC Press |
| Total Pages | : 432 |
| Release | : 2002-09-27 |
| Genre | : Science |
| ISBN | : 9780849300417 |
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 disciplines, including viral infections, gene modification, tumor diagnosis, gene therapy, and cellular distribution of rare mRNA copies. PCR/RT-PCR in situ: Light and Electron Microscopy covers methods of in situ polymerase chain reaction (PCR) and reverse transcription PCR (RT-PCR), two new approaches in visualizing very low amounts of DNA and RNA in tissues and cell cultures at the light and electron microscopy levels. Written by experts in this field, the book provides theoretical consideration, as well as practical approaches to in situ PCR. The authors provide detailed protocols for each step, including the preparation of tissue samples, the rationale for the design of primers and revelation. They also emphasize the need for appropriate controls to meet the requirements of in situ PCR and RT-PCR specificity. Organized in a user-friendly two-column format, this book will provide you with tools necessary to perform and optimize these sensitive and powerful techniques in your research protocols.
| Author | : C. S. Herrington |
| Publisher | : |
| Total Pages | : 228 |
| Release | : 1998 |
| Genre | : DNA replication |
| ISBN | : 9780199636327 |
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.
| Author | : Omar Bagasra |
| Publisher | : Wiley-Liss |
| Total Pages | : 0 |
| Release | : 1997-07-04 |
| Genre | : Science |
| ISBN | : 9780471159469 |
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.
| Author | : John M. S. Bartlett |
| Publisher | : Springer Science & Business Media |
| Total Pages | : 787 |
| Release | : 2008-02-02 |
| Genre | : Medical |
| ISBN | : 1592590713 |
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.
| Author | : Franck Pellestor |
| Publisher | : Springer Science & Business Media |
| Total Pages | : 257 |
| Release | : 2008-02-03 |
| Genre | : Science |
| ISBN | : 1597450685 |
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.
| Author | : Nicola King |
| Publisher | : Springer Science & Business Media |
| Total Pages | : 370 |
| Release | : 2008-02-04 |
| Genre | : Science |
| ISBN | : 159259283X |
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.
| Author | : Elizabeth van Pelt-Verkuil |
| Publisher | : Springer Science & Business Media |
| Total Pages | : 333 |
| Release | : 2008-03-14 |
| Genre | : Science |
| ISBN | : 1402062419 |
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).
| Author | : John M. S. Bartlett |
| Publisher | : Springer Science & Business Media |
| Total Pages | : 1083 |
| Release | : 2008-02-03 |
| Genre | : Science |
| ISBN | : 1592593844 |
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.
| Author | : Jiang Gu |
| Publisher | : Birkhäuser |
| Total Pages | : 143 |
| Release | : 2012-02-25 |
| Genre | : Science |
| ISBN | : 9781468468458 |
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.
