Development of New Analytical Method for Specific Nucleic Acid Sequences Using Affinity Capillary Electrophoresis PDF Download
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Author: 足立賢
Publisher:
ISBN:
Category :
Languages : en
Pages : 104
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Book Description
Author: 足立賢
Publisher:
ISBN:
Category :
Languages : en
Pages : 104
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Book Description
Author: Keith R. Mitchelson
Publisher: Springer Science & Business Media
ISBN: 1592590551
Category : Science
Languages : en
Pages : 481
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Book Description
The development of PCR, which enables extremely small amounts of DNA to be amplified, led to the rapid development of a multiplicity of a- lytical procedures that permit use of this new resource for the analysis of genetic variation and for the detection of disease-causing mutations. The advent of capillary electrophoresis (CE), with its power to separate and a- lyze very small amounts of DNA, has also stimulated researchers to develop analytical procedures for the CE format. The advantages of CE in terms of speed and reproducibility of analyses are manifold. Furthermore, the high s- sitivity of detection, and the ability to increase sample throughput with par- lel analysis, has led to the creation of a full range of analysis of DNA molecules, from modified DNA adducts and single-strand oligonucleotides through PCR-amplified DNA fragments and whole chromosomes. Capillary Elect- phoresis of Nucleic Acids focuses on analytical protocols that can be used for detection and analysis of mutations and modification, from precise DNA loci through entire genomes of organisms. Important practical considerations for CE, such as the choice of separation media, electrophoresis conditions, and the influence of buffer additives and dyes on DNA mobility, are discussed in several key chapters and within particular applications.
Author: Keith R. Mitchelson
Publisher: Humana Press
ISBN: 9781617371868
Category : Science
Languages : en
Pages : 484
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Book Description
The development of PCR, which enables extremely small amounts of DNA to be amplified, led to the rapid development of a multiplicity of a- lytical procedures that permit use of this new resource for the analysis of genetic variation and for the detection of disease-causing mutations. The advent of capillary electrophoresis (CE), with its power to separate and a- lyze very small amounts of DNA, has also stimulated researchers to develop analytical procedures for the CE format. The advantages of CE in terms of speed and reproducibility of analyses are manifold. Furthermore, the high s- sitivity of detection, and the ability to increase sample throughput with par- lel analysis, has led to the creation of a full range of analysis of DNA molecules, from modified DNA adducts and single-strand oligonucleotides through PCR-amplified DNA fragments and whole chromosomes. Capillary Elect- phoresis of Nucleic Acids focuses on analytical protocols that can be used for detection and analysis of mutations and modification, from precise DNA loci through entire genomes of organisms. Important practical considerations for CE, such as the choice of separation media, electrophoresis conditions, and the influence of buffer additives and dyes on DNA mobility, are discussed in several key chapters and within particular applications.
Author: Christoph Heller
Publisher: Springer Science & Business Media
ISBN: 3322910156
Category : Technology & Engineering
Languages : en
Pages : 318
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Book Description
Die Kapillarelektrophorese (CE) ist heute in biochemischen Laboratorien noch nicht als Routinemethode zur DNA-Trennung etabliert, obwohl sie diverse Vorzüge gegenüber den derzeitigen Methoden besitzt. In diesem Band stellen verschiedene Wissenschaftler die theoretischen Aspekte der Trennung von Oligonucleinsäuren mit der CE, die instrumentellen Grundlagen und deren Grenzen und schließlich die praktische Anwendung in der Biochemie und Molekularbiologie vor. This book on capillary electrophoresis is unique in its focus on the separation of nucleic acids. The importance of electrophoretic separation in every molecular biology laboratory justifies this specialization, which is also reflected in the development of instrumentation. This book is aimed to help to implement this rather new and promising technology in the biological laboratories and to help to overcome typical problems that can occur when starting with a new technique. It should also trigger further development in the field. It covers the theoretical background as well as practical examples of usual applications. The authors are all experts in their field, having many years of experience with capillary electrophoresis and their application to nucleic acids.
Author: Andras Guttman
Publisher: Newnes
ISBN: 0080931359
Category : Science
Languages : en
Pages : 391
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Book Description
Capillary Gel Electrophoresis and Related Microseparation Techniques covers all theoretical and practical aspects of capillary gel electrophoresis. It also provides an excellent overview of the key application areas of nucleic acid, protein and complex carbohydrate analysis, affinity-based methodologies, micropreparative aspects and related microseparation methods. It not only gives readers a better understanding of how to utilize this technology, but also provides insights into how to determine which method will provide the best technical solutions to particular problems. This book can also serve as a textbook for undergraduate and graduate courses in analytical chemistry, analytical biochemistry, molecular biology and biotechnology courses. - Covers all theoretical and practical aspects of capillary gel electrophoresis - Excellent overview of the key applications of nucleic acid, protein and complex carbohydrate analysis, affinity-based methodologies, micropreparative aspects and related microseparation methods - Teaches readers how to use the technology and select methods that are ideal for fundamental problems - Can serve as a textbook for undergraduate and graduate courses in analytical chemistry, analytical biochemistry, molecular biology and biotechnology courses
Author: Jennifer Lee Zabzdyr
Publisher:
ISBN:
Category : Capillary electrophoresis
Languages : en
Pages : 334
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Book Description
Author:
Publisher: Springer
ISBN: 9781592590551
Category : Capillary electrophoresis
Languages : en
Pages :
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Book Description
Author: Dietmar Tietz
Publisher: Boom Koninklijke Uitgevers
ISBN: 9783540639596
Category : Medical
Languages : en
Pages : 354
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Book Description
Electrophoresis is a powerful method to analyze nucleic acids (DNA, RNA). Various sophisticated techniques such as capillary electrophoresis, pulsed-field electrophoresis, fingerprinting using RFLP and RAPD, DNA sequencing, and mobility shift assay are described in detail. The required apparatus, appropriate use, preparation of probes, gel staining, interpretation of results, tricks for troubleshooting, manufacturers addresses, helpful Internet resources as well as specific applications, e.g. in legal medicine, microbiology and agriculture, are presented by leading experts.
Author:
Publisher:
ISBN:
Category : Medicine
Languages : en
Pages : 1064
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Book Description
Author: Alexandre Louis Andre Persat
Publisher:
ISBN:
Category :
Languages : en
Pages :
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Book Description
Microfluidics has recently enabled new capabilities in life sciences research. By leveraging physics at the microscale, novel miniaturized methods and devices provide fundamental improvements over traditional assays including higher sensitivity, massive parallelization, speed and automation. For example, nucleic acid analyses such as PCR or capillary electrophoresis are now commonly executed on microfluidic platforms. However, extracting and isolating a set of molecules of interest from a biological sample remains a widespread challenge, in particular when the target is a nucleic acid; such sample preparation has been identified as the "weak link" of microfluidics. Moreover, the discovery of classes of small RNA such as microRNA (miRNA) has revealed the limitations of benchtop preparation methods. This work tackles these issues by leveraging an electrophoretic focusing method, isotachophoresis (ITP), to perform selective focusing of specific nucleic acid molecules contained in complex mixtures. This includes extraction of genomic DNA from blood and isolation of miRNA from total RNA. Also, we leverage the unique physics of ITP to perform simultaneous purification and analysis of these molecules, thus enabling automated analysis at unprecedented speed. ITP is a robust electrophoretic preconcentration technique which generates strong electric field gradients, and enables selective focusing and separation of charged species based on their electrophoretic mobilities. In this work, we show that we can extract and purify genomic DNA from chemically lysed whole blood samples by carefully controlling ITP electrolyte chemistry to achieve selective focusing of genetic material. We show that ITP outputs PCR-compatible DNA with high efficiency in about 1 min. This novel sample preparation technique allows for efficient, fast, and automated purification of DNA from 10 nL to 1 [Mu]L of biological fluids. We also demonstrate that ITP focusing of microRNA -- short (~22 nt), non-coding RNA regulating gene expression -- enables quantification and sequence specific detection. We leverage both the selective and preconcentration capability of ITP for the quantification of global miRNA abundance. This allows for the measurement of RNA silencing activity in specific cells or tissues. We have optimized ITP chemistry and used a multi-stage injection strategy to selectively preconcentrate and quantify RNA shorter than 40 nt. We discuss results of miRNA quantification for a wide variety of samples. Additionally, we show that the combination of selective ITP focusing with simultaneous hybridization with molecular beacons is an efficient method for detection and quantitation of specific miRNA sequences. We demonstrate the efficacy of this assay for the detection of a liver specific miRNA. Finally, we show that we can use ITP to perform polymerase chain reaction in isothermal conditions by creating a cycles of chemicals mimicking thermal cycling.
