Multiplexing of Singleplex Real-Time-PCR Assays into Duplexes and Triplexes

Multiplexing of Singleplex Real-Time-PCR Assays into Duplexes and Triplexes

Author: Patrick Maaß

Publisher: diplom.de

Published: 2011-05-16

Total Pages: 58

ISBN-13: 3842814372

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Inhaltsangabe:Introduction: Breast Cancer: Cancer describes a group of various diseases, where cells start changing their molecular structure and begin to grow and to supersede normal cells. Cancer is induced by numerous different elicitors, which finally all lead to an interference of the genetically regulated balance between cell cycle and apoptosis. Although every organ in the human body can be afflicted with cancer, there are significant differences in frequency relating amongst others to age, sex, geographic region and personal habits. In the industrialized countries, breast cancer is the leading cause of death for women at the age between 30 and 60 years. With estimated 636.000 incident cases in the developed countries and 514.000 in the developing countries, breast cancer is the most prevalent cancer type among woman worldwide. Once detected, the cancer is classified based upon pathological characterizations of the tumor or a biopsy and the lymph nodes. A clinical way of characterizing the tumor is the TNM-classification, which describes the size of the tumor (T), the number of affected lymph nodes (N) and the existence of distant metastases (M). The histological classification characterizes the carcinoma according to its structural and cellular appearance and the amitosis rate leading to a grading from 1 to 3. An immuno-histological examination provides information about the estrogen- and the progesterone-receptor- and about the Her-2/neu-status. Breast cancer is a very heterogeneous disease. There are basic classifications that are unquestioned, even today. Recent studies confirmed the need to determine well known markers (i.e. estrogen (ER) and progesterone (PR) receptor status or HER2 status), but the large variety of subtypes and the corresponding different molecular pattern impede a uniform treatment. Although already today other factors than anatomical classifications are being taken into consideration, there exists a need for further biological markers to assist the physician in charge with his evaluation. Beside the diagnostic recognition, the choice of appropriate therapy and the prediction of prognosis are goals that should be reached in order to prevent early stage cancer patients from therapies that provide minimal benefit but reduce their quality of life by intense adverse reactions. RNA Expression Profiling and Prediction: Already today there are numerous genes associated with breast cancer occurrence, therapy [...]


Multiplexing of PCR Assays in Breast Cancer Analysis

Multiplexing of PCR Assays in Breast Cancer Analysis

Author: Patrick Maaß

Publisher: diplom.de

Published: 2012-03-01

Total Pages: 54

ISBN-13: 3863416074

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Das Werk beschreibt die Optimierung eines prognostischen molekularbiologischen Tests für die Brustkrebs-Diagnose und -Therapieentscheidung. Im Rahmen der Arbeit wurde die Zahl der Reaktionsräume für Polymerasekettenreaktionen (PCR) durch die Optimierung vom Multiplexen derart reduziert, dass parallele und kosteneffizientere Analysen möglich wurden.


Protocols in Advanced Genomics and Allied Techniques

Protocols in Advanced Genomics and Allied Techniques

Author: Aruna Pal

Publisher: Springer Nature

Published: 2021-11-14

Total Pages: 762

ISBN-13: 1071618180

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This laboratory manual includes the latest tools and techniques involved in genomic research. It starts with an introductory chapter on genomics and the various tools and applications involved. The initial chapters present protocols for basic techniques such as DNA isolation, electrophoresis, PCR, cDNA synthesis etc. The book then goes on to describe more advanced techniques such as next-generation sequencing, exome sequencing, use of RNAi, RNAseq, genome editing, single cell genomics etc. Each topic includes a brief description, information on the principles involved, materials & methods, protocol, and expected results, with diagrams and graphs. All protocols are presented in a very lucid and precise way, to make it easy for readers to follow and replicate them.


Testing and Analysis of GMO-containing Foods and Feed

Testing and Analysis of GMO-containing Foods and Feed

Author: Salah E. O. Mahgoub

Publisher: CRC Press

Published: 2019-01-15

Total Pages: 305

ISBN-13: 135171385X

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An increasing number of genetically modified organisms (GMOs) continues to be produced every day. In response to the concerns raised by the development of GMOs and their incorporation in foods and feed, guidelines and regulations to govern and control the use of GMOs and their products have been enacted. These regulations necessitated the design of methods to detect and analyse the presence of GMOs or their products in agriculture produce, food and feed production chains. Design of techniques and instruments that would detect, identify, and quantify GM ingredients in food and feed will help inspection authorities to relay reliable information to consumers who might be concerned about the presence of GM ingredients. Information generated by detection of GMOs in food and feed would be helpful for setting regulations that govern the use of GM components as well as for labeling purposes. Qualitative detection methods of GM-DNA sequences in foods and feeds have evolved fast during the past few years. There is continuous need for the development of more advanced multi-detection systems and for periodic updates of the databases related to these systems. Testing and Analysis of GMO-containing Foods and Feed presents updates and comprehensive views on the various methods and techniques in use today for the detection, identification and quantification of GMOs in foods and feed. The eleven book chapters cover recent developments on sample preparation techniques, immunoassays methods and the PCR technique used in GMO analysis, the use of biosensors in relation to GMO analysis, the application of nucleic acid microarrays for the detection of GMOs, validation and standardization methods for GMO testing, in addition to the type of reference material and reference methods used in GMO testing and analysis. Some of the ISO standards designed for identifying and detecting the presence of GM material in foods are also presented in the book.


Gene Quantification

Gene Quantification

Author: Francois Ferre

Publisher: Springer Science & Business Media

Published: 2012-12-06

Total Pages: 379

ISBN-13: 1461241642

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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.


MoneyBall Medicine

MoneyBall Medicine

Author: Harry Glorikian

Publisher: Taylor & Francis

Published: 2017-11-20

Total Pages: 591

ISBN-13: 1351984330

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How can a smartwatch help patients with diabetes manage their disease? Why can’t patients find out prices for surgeries and other procedures before they happen? How can researchers speed up the decade-long process of drug development? How will "Precision Medicine" impact patient care outside of cancer? What can doctors, hospitals, and health systems do to ensure they are maximizing high-value care? How can healthcare entrepreneurs find success in this data-driven market? A revolution is transforming the $10 trillion healthcare landscape, promising greater transparency, improved efficiency, and new ways of delivering care. This new landscape presents tremendous opportunity for those who are ready to embrace the data-driven reality. Having the right data and knowing how to use it will be the key to success in the healthcare market in the future. We are already starting to see the impacts in drug development, precision medicine, and how patients with rare diseases are diagnosed and treated. Startups are launched every week to fill an unmet need and address the current problems in the healthcare system. Digital devices and artificial intelligence are helping doctors do their jobs faster and with more accuracy. MoneyBall Medicine: Thriving in the New Data-Driven Healthcare Market, which includes interviews with dozens of healthcare leaders, describes the business challenges and opportunities arising for those working in one of the most vibrant sectors of the world’s economy. Doctors, hospital administrators, health information technology directors, and entrepreneurs need to adapt to the changes effecting healthcare today in order to succeed in the new, cost-conscious and value-based environment of the future. The authors map out many of the changes taking place, describe how they are impacting everyone from patients to researchers to insurers, and outline some predictions for the healthcare industry in the years to come.


Pine Wilt Disease

Pine Wilt Disease

Author: Bo Guang Zhao

Publisher: Springer Science & Business Media

Published: 2008-09-27

Total Pages: 486

ISBN-13: 4431756558

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Pine forests face a global threat of pine wilt disease, which is being spread by vector beetles carrying pathogenic nematodes from dead trees to healthy ones. Among the host pines there are varying degrees of susceptibility, and nematode strains also contain a variety of virulences, both of which factors help to determine whether infected host trees will die or survive. As well, biotic and abiotic environmental factors influence the fate of infected trees. This book describes the history of the disease, pathogenic nematodes, vector beetles, the etiology and ecology of the disease, microorganisms involved, and control methods that utilize host resistance and biological control agents. Concrete, comprehensive, and the most up-to-date knowledge about this worldwide forest epidemic is presented for readers, enabling them to understand the nature and epidemic threat of pine wilt disease.


Molecular Microbiology

Molecular Microbiology

Author: David H. Persing

Publisher: John Wiley & Sons

Published: 2020-07-24

Total Pages: 852

ISBN-13: 1555819079

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Presenting the latest molecular diagnostic techniques in one comprehensive volume The molecular diagnostics landscape has changed dramatically since the last edition of Molecular Microbiology: Diagnostic Principles and Practice in 2011. With the spread of molecular testing and the development of new technologies and their opportunities, laboratory professionals and physicians more than ever need a resource to help them navigate this rapidly evolving field. Editors David Persing and Fred Tenover have brought together a team of experienced researchers and diagnosticians to update this third edition comprehensively, to present the latest developments in molecular diagnostics in the support of clinical care and of basic and clinical research, including next-generation sequencing and whole-genome analysis. These updates are provided in an easy-to-read format and supported by a broad range of practical advice, such as determining the appropriate type and quantity of a specimen, releasing and concentrating the targets, and eliminating inhibitors. Molecular Microbiology: Diagnostic Principles and Practice Presents the latest basic scientific theory underlying molecular diagnostics Offers tested and proven applications of molecular diagnostics for the diagnosis of infectious diseases, including point-of-care testing Illustrates and summarizes key concepts and techniques with detailed figures and tables Discusses emerging technologies, including the use of molecular typing methods for real-time tracking of infectious outbreaks and antibiotic resistance Advises on the latest quality control and quality assurance measures Explores the increasing opportunities and capabilities of information technology Molecular Microbiology: Diagnostic Principles and Practice is a textbook for molecular diagnostics courses that can also be used by anyone involved with diagnostic test selection and interpretation. It is also a useful reference for laboratories and as a continuing education resource for physicians. If you are looking for online access to the latest clinical microbiology content, please visit www.wiley.com/learn/clinmicronow.