PCR 3

PCR 3
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.

PCR for Clinical Microbiology

PCR for Clinical Microbiology
Author: Ian W.J. Carter
Publisher: Springer Science & Business Media
Total Pages: 420
Release: 2010-07-03
Genre: Science
ISBN: 9048190398

Not another textbook, but a valuable tool for doctors and microbiologists wanting to know how to set up a PCR diagnostic microbiology laboratory according to current regulatory standards and perform assays supplied with patient clinical diagnostic criteria and easy to follow protocols. Whether laboratories are using commercial kits or in-house methods developed in their own laboratories or adopted from published methods, all clinical microbiology laboratories need to be able to understand, critically evaluate, perform and interpret these tests according to rigorous and clinically appropriate standards and international guidelines. The cost and effort of development and evaluation of in-house tests is considerable and many laboratories do not have the resources to do so. This compendium is a vehicle to improve and maintain the clinical relevance and high quality of diagnostic PCR. It is a unique collection of; guidelines for PCR laboratory set up and quality control, test selection criteria, methods and detailed step by step protocols for a diagnostic assays in the field of molecular microbiology. The structure of the book provides the PCR fundamentals and describes the clinical aspects and diagnosis of infectious disease. This is followed by protocols divided into; bacteria, virus, fungi and parasites, and susceptibility screens. The inclusion of medical criteria and interpretation adds value to the compendium and benefits clinicians, scientists, researchers and students of clinical diagnostic microbiology

PCR Technology

PCR Technology
Author: Tania Nolan
Publisher: CRC Press
Total Pages: 461
Release: 2013-06-13
Genre: Law
ISBN: 143984805X

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.

PCR Protocols

PCR Protocols
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.

Pennsylvania Corporation Reporter

Pennsylvania Corporation Reporter
Author: George Ross Hull
Publisher:
Total Pages: 816
Release: 1916
Genre: Corporation law
ISBN:

Containing opinions, general orders, administrative rulings, reports, circulars, rules of practice, rules and regulations of service, etc., of the Public Service Commission of Pennsylvania; and opinions of the county courts throughout the Commonwealth and of the attorney general involving the law of private corporations, including corporation tax cases and appeals from the Public Service Commission; and index and annotations to the Public service company law.

Introduction to Plant Biotechnology (3/e)

Introduction to Plant Biotechnology (3/e)
Author: H S Chawla
Publisher: CRC Press
Total Pages: 745
Release: 2011-05-24
Genre: Science
ISBN: 1439894140

This book has been written to meet the needs of students for biotechnology courses at various levels of undergraduate and graduate studies. This book covers all the important aspects of plant tissue culture viz. nutrition media, micropropagation, organ culture, cell suspension culture, haploid culture, protoplast isolation and fusion, secondary metabolite production, somaclonal variation and cryopreservation. For good understanding of recombinant DNA technology, chapters on genetic material, organization of DNA in the genome and basic techniques involved in recombinant DNA technology have been added. Different aspects on rDNA technology covered gene cloning, isolation of plant genes, transposons and gene tagging, in vitro mutagenesis, PCR, molecular markers and marker assisted selection, gene transfer methods, chloroplast and mitochondrion DNA transformation, genomics and bioinformatics. Genomics covers functional and structural genomics, proteomics, metabolomics, sequencing status of different organisms and DNA chip technology. Application of biotechnology has been discussed as transgenics in crop improvement and impact of recombinant DNA technology mainly in relation to biotech crops.

PCR

PCR
Author: Mike McPherson
Publisher: Garland Science
Total Pages: 292
Release: 2007-01-25
Genre: Science
ISBN: 0203002679

A thoroughly updated version of the successful first edition with a new chapter on Real-Time PCR, more prokaryotic applications, and more detail in the complex mutagenesis sections. Information on PCR applications in genomics and proteomics have been expanded and integrated throughout the text. There is also advice on available products and specific pointers to the most appropriate methods. As with the first edition, this will be an ideal practical introduction and invaluable guide to PCR and its applications.

PCR Cloning Protocols

PCR Cloning Protocols
Author: Bing-Yuan Chen
Publisher: Springer Science & Business Media
Total Pages: 429
Release: 2008-02-05
Genre: Science
ISBN: 1592591779

PCR Cloning Protocols, Second Edition, updates and expands Bruce White's best-selling PCR Cloning Protocols (1997) with the newest procedures for DNA cloning and mutagenesis. Here the researcher will find readily reproducible methods for all the major aspects of PCR use, including PCR optimization, computer programs for PCR primer design and analysis, and novel variations for cloning genes of special characteristics or origin, with emphasis on long distance PCR and GC-rich template amplification. Also included are both conventional and novel enzyme-free and restriction site-free procedures to clone PCR products into a range of vectors, as well as state-of-the-art protocols to facilitate DNA mutagenesis and recombination, and to clone the challenging uncharacterized DNA flanking a known DNA fragment.

RT-PCR Protocols

RT-PCR Protocols
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.