Download or read book Quantitation of mRNA by Polymerase Chain Reaction written by Thomas Köhler and published by Springer Science & Business Media. This book was released on 2012-12-06 with total page 178 pages. Available in PDF, EPUB and Kindle. Book excerpt: In this laboratory "cook-book", the authors provide a concise guide to PCR-based techniques to quantify nucleic acids in biological and clinical samples using exclusively nonradioactive detection methods, e.g. HPLC, biotin and digoxigenin based protocols. Each method presentation also includes sections on theory, reagents, standards, applicability, limitations, and trouble shooting. In addition to the protocols, the authors also provide the necessary information on: general aspects of nucleic acid quantitation; design of PCR standards; mRNA purification; cDNA synthesis; solution hybridization; DNA sequencing. This laboratory guide enables professionals as well as beginners to adopt easily quantitative PCR protocols into their own clinical or biomedical research.
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 Rapid Cycle Real Time PCR written by S. Meuer and published by Springer Science & Business Media. This book was released on 2012-12-06 with total page 390 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.
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 Basic Science Techniques in Clinical Practice written by Hitendra R.H. Patel and published by Springer Science & Business Media. This book was released on 2008-08-25 with total page 162 pages. Available in PDF, EPUB and Kindle. Book excerpt: A complete guide to implementing research projects for anyone in the medical professions. This book covers all the main areas, allowing anyone to set up and complete research projects. The techniques outlined here can easily be adapted to clinical projects. Written by international authors to provide a flavor from many institutions, the book’s appeal is cross-sectional, both at hospital and primary care levels worldwide. Providing cutting-edge information in an accessible manner, and containing diagrams and easy-to-follow step-by-step guides, this is the first guide of its kind. It contains a complete section on setting up and funding research projects.
Download or read book Calibrated Reverse Transcriptase polymerase Chain Reaction for Quantitation of MRNA written by Mustafa Vakur Bor and published by . This book was released on 2000 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:
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 PCR Topics written by Arndt Rolfs and published by Springer Science & Business Media. This book was released on 2013-12-01 with total page 258 pages. Available in PDF, EPUB and Kindle. Book excerpt: PCR, developed at Cetus Corporation/USA by Henry A. Erlich, Kary Mullis and Randall K. Saiki, is a very simple method for amplifying nucleic acids in vitro. The realization of this idea bases on the repetition of a set of three different temperatures and yields an increase of the target structure up to a factor of 106 to 1012. Therefore, this technique is predisposed for safe analysis and characterization of DNA and RNA sequences of interest, even where the starting amount of material is enormously small. Because of its sensitivity, speed and versatility this method is particularly suitable for investigations of oncogenes, tumor associated translocations, retroviral sequences, lymphokines and mainly the broad field of degenerative and inflammatory diseases of nervous system. PCR seems to be the technique which could overcome the two most important problems in that field: very small amount of material combined with the necessity of rapid diagnostic procedures in inflammatory infections. "PCR topics" will give an actual overview of basic and applied research fields on usage of polymerase chain reaction. All contributions to this book have been presented at an international congress on "Usage of Polymerase chain reaction in genetic and infectious diseases" which took place in june 1990 in Berlin. The editors wish to thank all participants for their contributions. We offer our thanks and gratitude to our coworkers and especially to our technical assistents Barbara Trampenau, Mirjana Wiirdemann and Hannelore Leonhard.
Download or read book RNA Based Regulation in Human Health and Disease written by and published by Academic Press. This book was released on 2020-08-19 with total page 444 pages. Available in PDF, EPUB and Kindle. Book excerpt: RNA-based Regulation in Human Health and Disease offers an in-depth exploration of RNA mediated genome regulation at different hierarchies. Beginning with multitude of canonical and non-canonical RNA populations, especially noncoding RNA in human physiology and evolution, further sections examine the various classes of RNAs (from small to large noncoding and extracellular RNAs), functional categories of RNA regulation (RNA-binding proteins, alternative splicing, RNA editing, antisense transcripts and RNA G-quadruplexes), dynamic aspects of RNA regulation modulating physiological homeostasis (aging), role of RNA beyond humans, tools and technologies for RNA research (wet lab and computational) and future prospects for RNA-based diagnostics and therapeutics. One of the core strengths of the book includes spectrum of disease-specific chapters from experts in the field highlighting RNA-based regulation in metabolic & neurodegenerative disorders, cancer, inflammatory disease, viral and bacterial infections. We hope the book helps researchers, students and clinicians appreciate the role of RNA-based regulation in genome regulation, aiding the development of useful biomarkers for prognosis, diagnosis, and novel RNA-based therapeutics. Comprehensive information of non-canonical RNA-based genome regulation modulating human health and disease Defines RNA classes with special emphasis on unexplored world of noncoding RNA at different hierarchies Disease specific role of RNA - causal, prognostic, diagnostic and therapeutic Features contributions from leading experts in the field
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 258 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.
Download or read book Polymerase Chain Reaction for Biomedical Applications written by Ali Samadikuchaksaraei and published by BoD – Books on Demand. This book was released on 2016-12-14 with total page 186 pages. Available in PDF, EPUB and Kindle. Book excerpt: Do you want to know the details that should be taken into consideration in order to have accurate conventional and real-time PCR results? If so, this book is for you. Polymerase Chain Reaction for Biomedical Applications is a collection of chapters for both novice and experienced scientists and technologists aiming to address obtaining an optimized real-time PCR result, simultaneous processing of a large number of samples and assays, performing PCR and RT-PCR on cell lysate without extraction of DNA or RNA, detecting false-positive PCR results, detecting organisms in viral and microbial diseases and hospital environment, following safety assessments of food products, and using PCR for introduction of mutations. This is a must-have book for any PCR laboratory.
Download or read book The Polymerase Chain Reaction written by Kary B. Mullis and published by Springer Science & Business Media. This book was released on 2012-02-02 with total page 464 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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, " ...
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 The Polymerase Chain Reaction written by Kary B. Mullis and published by Springer Science & Business Media. This book was released on 1994 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: One: Methodology.- I. Basic Methodology.- 1. Manipulation of DNA by PCR.- 2. Cloning PCR Products.- 3. Optimization of Multiplex PCRs.- 4. Preparation of Nucleic Acids for Archival Material.- 5. PCR Amplification of Viral DNA and Viral Host Cell mRNAs in Situ.- II. Quantitation.- 6. Quantitative PCR: An Overview.- 7. Quantification of DNAs by the Polymerase Chain Reaction Using an Internal Control.- 8. RT-PCR and mRNA Quantitation.- 9. Analysis of Human T-Cell Repertoires by PCR.- III. Nonisotopic Detection.- 10. Ultrasensitive Nonradioactive Detection of PCR Reactions: An Overview.- 11. Fluorescent Detection Methods for PCR Analysis.- 12. Enzyme-Labeled Oligonucleotides.- 13. Application of the Hybridization Protection Assay (HPA) to PCR.- IV. Instrumentation.- 14. PCR Instrumentation: Where Do We Stand?.- 15. Rapid Cycle DNA Amplification.- 16. Automating the PCR Process.- V. Sequencing.- 17. PCR and DNA Sequencing.- 18. Phage Promoter-Based Methods for Sequencing and Screening for Mutations.- 19. Capture PCR: An Efficient Method for Walking Along Chromosomal DNA and cDNA.- Two: Applications.- I. General Applications.- 20. In Vitro Evolution of Functional Nucleic Acids: High-Affinity RNA Ligands of the HIV-1 rev Protein.- 21. The Application of PCR to Forensic Science.- 22. Recreating the Past by PCR.- 23. Nonbiological Applications.- II. Genetic Analysis.- 24. RT-PCR and Gene Expression.- 25. Fingerprinting Using Arbitrarily Primed PCR: Application to Genetic Mapping, Population Biology, Epidemiology, and Detection of Differentially Expressed RNAs.- 26. Genetics, Plants, and the Polymerase Chain Reaction.- III. Assessment of Therapy Effectiveness.- 27. PCR Assessment of the Efficacy of Therapy in Philadelphia Chromosome-Positive Leukemias.- 28. The Detection of Minimal Residual Disease (MRD) in Acute Lymphoblastic Leukemia Using Clone-Specific Probes Directed against V(D)J Junctional Sequences.- 29. Assessment of Therapy Effectiveness: Infectious Disease.- 30. Gene Therapy.- IV. Diagnostics.- 31. PCR and Cancer Diagnostics: Detection and Characterization of Single Point Mutations in Oncogenes and Antioncogenes.- 32. Clinical Applications of the Polymerase Chain Reaction.- 33. Infectious Diseases.- Three: PCR and the World of Business.- 34. PCR in the Marketplace.- 35. PCR and Scientific Invention: The Trial of DuPont vs. Cetus.
Download or read book Real time PCR written by M Tevfik Dorak and published by Garland Science. This book was released on 2007-02-08 with total page 333 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 The PCR Revolution written by Stephen A. Bustin and published by Cambridge University Press. This book was released on 2010 with total page 327 pages. Available in PDF, EPUB and Kindle. Book excerpt: Examines the latest innovations and the overall impact of PCR on areas of molecular research.
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.
