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Author: S. Meuer
Publisher: Springer Science & Business Media
ISBN: 3642595243
Category : Science
Languages : en
Pages : 390
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Book Description
The first comprehensive treatise on Rapid Cycle Real-Time PCR. With amplification times of 15-30 minutes of on-line detection and analysis, nucleic acid quantification of mutation analysis finally becomes a routine, powerful and rapid method. Focusing primarily on the LightCycler, an instrument that combines Rapid Cycle PCR with fluorescent monitoring, this technology provides convenient analysis by melting temperatures. PCR products can be identified by product Tm, and single base mismatches can easily be genotyped by probe Tm. Methods chapters detail the theory behind quantification of mutation analysis; the design of synthesis of fluorescent hybridization probes of the preparation of template DNA. Application chapters apply nucleid acid quantification to infectious organisms of intracellular messengers and mutation detection to somatic of acquired mutations.
Author: S. Meuer
Publisher: Springer Science & Business Media
ISBN: 3642595243
Category : Science
Languages : en
Pages : 390
Get Book Here
Book Description
The first comprehensive treatise on Rapid Cycle Real-Time PCR. With amplification times of 15-30 minutes of on-line detection and analysis, nucleic acid quantification of mutation analysis finally becomes a routine, powerful and rapid method. Focusing primarily on the LightCycler, an instrument that combines Rapid Cycle PCR with fluorescent monitoring, this technology provides convenient analysis by melting temperatures. PCR products can be identified by product Tm, and single base mismatches can easily be genotyped by probe Tm. Methods chapters detail the theory behind quantification of mutation analysis; the design of synthesis of fluorescent hybridization probes of the preparation of template DNA. Application chapters apply nucleid acid quantification to infectious organisms of intracellular messengers and mutation detection to somatic of acquired mutations.
Author: Carl Wittwer
Publisher: Springer
ISBN: 3642188400
Category : Science
Languages : en
Pages : 219
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Book Description
Rapid Cycle Real-Time PCR is a powerful technique for nucleic acid quantification and analysis that takes less than 30 minutes to complete. Fluorescence is automatically monitored each cycle and the amount of template quantified by advanced analytical methods, such as the second derivative maximum method. Immediately following rapid cycle PCR, melting curve analysis is performed to verify product purity with SYBR Green I and/or genotype with fluorescently-labeled hybridization probes(HybProbes or SimpleProbes). Rapid cycle real-time PCR is often cited as the most versatile, efficient method for nucleic acid quantification in research and climical studies. Molecular analysis has never been easier!
Author: Francois Ferre
Publisher: Springer Science & Business Media
ISBN: 1461241642
Category : Medical
Languages : en
Pages : 379
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Book Description
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.
Author: Kary B. Mullis
Publisher: Springer Science & Business Media
ISBN: 1461202574
Category : Medical
Languages : en
Pages : 464
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Book Description
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, " ...
Author: Kirstin J. Edwards
Publisher: Taylor & Francis
ISBN: 113418400X
Category : Polymerase chain reaction
Languages : en
Pages : 362
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Book Description
Author: Stephen A. Bustin
Publisher: Cambridge University Press
ISBN: 0521882311
Category : Science
Languages : en
Pages : 327
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Book Description
Examines the latest innovations and the overall impact of PCR on areas of molecular research.
Author: M Dorak
Publisher: Garland Science
ISBN: 1134183992
Category : Medical
Languages : en
Pages : 484
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Book Description
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.
Author: Carl Wittwer
Publisher: Springer Science & Business Media
ISBN: 9783540206293
Category : Medical
Languages : en
Pages : 240
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Book Description
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 allow 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. The method is often cited as the most versatile, efficient method for nucleic acid analysis in research and analysis in the fields of Genetics and Oncology. Molecular analysis has never been easier! TOC: Methods 1 Onno Bakker, Academic Medical Centre Amsterdam, NL Housekeeping Genes: A Gold Standard? 2 Weisser/ Schnittger, Klinikum Grosshadern München, Germany The choice of house keeping genes in MRD-quantification of t(8;21) positive AML 3 Ronald H. Lekanne-Deprez, Dep of Anatomie & Embryologie, Amsterdam, The Netherlands Quantification of mRNA Using Linear Regression of Log- Linear PCR Data-Points as an Alternative for the Standard Curve Approach 4 Jochen Wilhelm, University Giessen Estimation of Genome Sizes by Quantitative Real-Time PCR Applications Regulation and Development 5 N. Neubauer, University of Copenhagen, Biokemisk Afd., Copenhagen, Denmark Relative Quantification of Insulin Gene Expression on the LightCycler Using SYBR Green I 6 Jürgen Loeffler, Medizinische Klinik, Abt. II, Otfried-Müller-Str. 10, 72076 Tübingen, Germany Quantification of T-Cell Receptor Excision Circle DANN Using Fluorescence Resonance Energy Transfer and the LightCycler System 7 Jim Whelan, Plant Molecular Biology Group, University of Western Australia, Crawley, Australia Investigation of Mitochondrial Biogenesis in Plants using Quantitative Real-Time PCR 8 E. Veistinen, Turku University, Dept. Medical Microbiology, Kiinamyllynkatu 13, FIN 20520 Turku Quantification of Ikaros Family Isoforms by Real-Time PCR 9 P. Stordeur, Dep. Immunologie- Hematologie-Transfusion, Hopital Erasme, Brussels, Belgium Methods to quantify cytokine gene expression by Real-Time PCR Oncology 10 Dr. Bernard, Idahotech, Salt Lake City, USA quantitative profiling for breast cancer using DNA and RNA markers 11 Melanie Königshoff, University Giessen Quantification of HER-2/NEU Gene Copy Number in Breast Cancer Tissue 12 Remedios Castelló Cros, Dpto. Bioquímica. Centro de Investigación, Hospital la Fe, Av. Campanar, 21, 46009 Valencia, Spain Quantitative real-time reverse transcription-PCR assay for urokinase plasminogen activator, plasminogen activator inhibitor type 1, and tissue metalloproteinase inhibitor type 1 gene expressions in primary breast cancer 13 C. H. W. Klaasen, C. Wilhelmina Hospital, Dep. Of Med. Microbiology & Infectious Diseases Nijmegen, NL Relative Quantification of Human DNA in Feaces (stool) 14 Chung-Che (Jeff) Chang, Assistant Professor, Director, Hematopathology Fellowship and Molecular/Pharmacogenetics Lab., Dep of Pathology, Medical College of Wisconsin, 9200 W. Wisconsin Ave., Milwaukee, WI 53226 real- time quatification of tumor load (t(14;18))in follicular lymphoma patients 15 P. Bolufer, Laboratorio de Biologia Molecular, Universitario La Fe, Valencia, Spain Real time quantification of AML rearrangements (AML1/ETO and TEL/AML1 ) in the diagnosis and monitoring of acute leukemia Genetics 16 Francisco Barros, INGO, Santiago de Compostela Gene Dosage Determination by Real Time PCR 17 Elaine Lyon, ARUP Laboratories, Salt Lake City, USA deletions and duplications of the cytochrome p450 2D6 gene using a reference gene and competitor (Alison Millson) 18 Karin Berg, Pathology, John Hopkins Medical Inst, Baltimore, USA Analysis of Bone Marrow Engraftment Following in Utero Bone Marrow Transplantation in a Can
Author: Sean R. Gallagher
Publisher: John Wiley & Sons
ISBN: 047094241X
Category : Science
Languages : en
Pages : 679
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Book Description
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.
Author: U. Reischl
Publisher: Springer Science & Business Media
ISBN: 3642483518
Category : Science
Languages : en
Pages : 253
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Book Description
Rapid Cycle Real-Time PCR is a powerful analytical tool with broad application for the basic and applied life sciences. Compared with conventional PCR technology, Rapid Cycle Real-Time PCR is faster, has greater specificity, and is more easily adaptable for a variety of diagnostic tests, including qualitative, quantitative and mutation detection assays. This book provides general overviews of this technology for use in the clinical microbiology laboratory as well as specific diagnostic protocols for the detection of viral, bacterial and fungal pathogens and genetically modified organisms in human specimens and foodstuffs. All of these protocols have been developed, verified, and validated by experts in the field and should be of great interest for clinical microbiologists, pathologists, laboratory technologists as well as practicing physicians.
