Methods for Parallel Amplification of Single DNA Molecules PDF Download
Are you looking for read ebook online? Search for your book and save it on your Kindle device, PC, phones or tablets. Download Methods for Parallel Amplification of Single DNA Molecules PDF full book. Access full book title Methods for Parallel Amplification of Single DNA Molecules by Erin Marie McElfish. Download full books in PDF and EPUB format.
Author: Erin Marie McElfish Publisher: ISBN: Category : Languages : en Pages : 77
Book Description
The aim of this thesis is to develop two improved methods for amplifying genomic DNA and to order the individual amplified templates into arrays for high throughput and cost-efficient genome sequencing. The first method focuses on the synthesis of functionalized linear polymers for fabricating arrays for DNA amplification. These arrays can be used to improve PCR amplification of single molecules by extending the reaction into three-dimensional space similar to a solution reaction while each reaction is confined within the structure of the array. A method was developed for the synthesis and purification of linear polyacrylamide copolymers that can be captured and visualized. Several conjugation methods were investigated for functionalizing the copolymers with primers. The second amplification method utilized linear rolling circle amplification to produce long continuous DNA molecules with multiple copies of the template sequence. These single molecules could be captured onto an array for sequencing. The number of amplified copies in these molecules was characterized by several methods including gel electrophoresis, digestion, probe saturation, and electric field stretching. The amplified products displayed a broad distribution in length. However, they could be captured on a surface and imaged by fluorescence microscopy, and are accessible for downstream applications. Microemulsions can be used to further control the sizes of DNA single molecule amplicons. A microfluidic device was designed and fabricated for creating uniform microemulsions. Microemulsions with a coefficient of variation around 0.15 could be produced under various conditions. Further improvement in the device will be required to make it more reproducible.
Author: Erin Marie McElfish Publisher: ISBN: Category : Languages : en Pages : 77
Book Description
The aim of this thesis is to develop two improved methods for amplifying genomic DNA and to order the individual amplified templates into arrays for high throughput and cost-efficient genome sequencing. The first method focuses on the synthesis of functionalized linear polymers for fabricating arrays for DNA amplification. These arrays can be used to improve PCR amplification of single molecules by extending the reaction into three-dimensional space similar to a solution reaction while each reaction is confined within the structure of the array. A method was developed for the synthesis and purification of linear polyacrylamide copolymers that can be captured and visualized. Several conjugation methods were investigated for functionalizing the copolymers with primers. The second amplification method utilized linear rolling circle amplification to produce long continuous DNA molecules with multiple copies of the template sequence. These single molecules could be captured onto an array for sequencing. The number of amplified copies in these molecules was characterized by several methods including gel electrophoresis, digestion, probe saturation, and electric field stretching. The amplified products displayed a broad distribution in length. However, they could be captured on a surface and imaged by fluorescence microscopy, and are accessible for downstream applications. Microemulsions can be used to further control the sizes of DNA single molecule amplicons. A microfluidic device was designed and fabricated for creating uniform microemulsions. Microemulsions with a coefficient of variation around 0.15 could be produced under various conditions. Further improvement in the device will be required to make it more reproducible.
Author: Keith R. Mitchelson Publisher: Elsevier ISBN: 0080471285 Category : Science Languages : en Pages : 399
Book Description
Since the independent invention of DNA sequencing by Sanger and by Gilbert 30 years ago, it has grown from a small scale technique capable of reading several kilobase-pair of sequence per day into today's multibillion dollar industry. This growth has spurred the development of new sequencing technologies that do not involve either electrophoresis or Sanger sequencing chemistries. Sequencing by Synthesis (SBS) involves multiple parallel micro-sequencing addition events occurring on a surface, where data from each round is detected by imaging. New High Throughput Technologies for DNA Sequencing and Genomics is the second volume in the Perspectives in Bioanalysis series, which looks at the electroanalytical chemistry of nucleic acids and proteins, development of electrochemical sensors and their application in biomedicine and in the new fields of genomics and proteomics. The authors have expertly formatted the information for a wide variety of readers, including new developments that will inspire students and young scientists to create new tools for science and medicine in the 21st century. Reviews of complementary developments in Sanger and SBS sequencing chemistries, capillary electrophoresis and microdevice integration, MS sequencing and applications set the framework for the book. * 'Hot Topic' with DNA sequencing continuing as a major research activity in many areas of life science and medicine. * Bringing together new developments in DNA sequencing technology * Reviewing issues relevant to the new applications used
Author: Ulrich Finckh Publisher: Springer Science & Business Media ISBN: 1461525306 Category : Medical Languages : en Pages : 315
Book Description
The polymerase chain reaction (PCR) - an in Vitro techniques for producing large amounts of a specific DNA fragment - has rapidly become established as one of the most important, impressive and fascinating methods of molecular biology as well as clinical diagnostics. In the seven years since'the technique was published, it has had a major impact on medical research. However, as there are still problems in instruments, standardized protocols for diagnostic applications and unsolved difficulties to avoid cross-contaminations on the one hand and on the other hand the even present question of how to interpret the biological value of a PCR result, most clinicians prefer to further wait until these topics are clarified. It is the aim of this book to give the reader lab-proven protocols from experienced scientists as well as a general introduction to alternative DNA-amplification procedures and their possible usage such as the NASBA or LCR. This book is divided into four major parts to provide a theoretical (first and second section) and a practical framework for a better understanding of the new technology. In the first part we provide an up-to-date summary of basic problems in this rapidly evolving field. We demonstrate, for example how to use fixed tissue materials and how to quantify PCR products as well as how to prepare nucleic acids in a safe, convenient and proper way, or even how to sequence directly PCR products for the analysis of the DNA structure.
Author: Klaus D. Sattler Publisher: CRC Press ISBN: 1420075411 Category : Science Languages : en Pages : 816
Book Description
Covering the key theories, tools, and techniques of this dynamic field, Handbook of Nanophysics: Principles and Methods elucidates the general theoretical principles and measurements of nanoscale systems. Each peer-reviewed chapter contains a broad-based introduction and enhances understanding of the state-of-the-art scientific content through fund
Author: Tom Strachan Publisher: Garland Science ISBN: 1136844074 Category : Science Languages : en Pages : 808
Book Description
Human Molecular Genetics is an established and class-proven textbook for upper-level undergraduates and graduate students which provides an authoritative and integrated approach to the molecular aspects of human genetics. While maintaining the hallmark features of previous editions, the Fourth Edition has been completely updated. It includes new Key Concepts at the beginning of each chapter and annotated further reading at the conclusion of each chapter, to help readers navigate the wealth of information in this subject. The text has been restructured so genomic technologies are integrated throughout, and next generation sequencing is included. Genetic testing, screening, approaches to therapy, personalized medicine, and disease models have been brought together in one section. Coverage of cell biology including stem cells and cell therapy, studying gene function and structure, comparative genomics, model organisms, noncoding RNAs and their functions, and epigenetics have all been expanded.
Author: Shu Chien Publisher: World Scientific ISBN: 981270793X Category : Science Languages : en Pages : 563
Book Description
This bestselling textbook will introduce undergraduate bioengineering students to the fundamental concepts and techniques, with the basic theme of integrative bioengineering. It covers bioengineering of several body systems, organs, tissues, and cells, integrating physiology at these levels with engineering concepts and approaches; novel developments in tissue engineering, regenerative medicine, nanoscience and nanotechnology; state-of-the-art knowledge in systems biology and bioinformatics; and socio-economic aspects of bioengineering.One of the distinctive features of the book is that it is integrative in nature (integration of biology, medicine and engineering, across different levels of the biological hierarchy, and basic knowledge with applications). It is unique in that it covers fundamental aspects of bioengineering, cutting-edge frontiers, and practical applications, as well as perspectives of bioengineering development. Furthermore, it covers important socio-economical aspects of bioengineering such as ethics and entrepreneurism.
Author: Brian K. Nunnally Publisher: CRC Press ISBN: 1420029088 Category : Medical Languages : en Pages : 252
Book Description
While one marvels at the DNA generated by the Human Genome Project, perhaps just as significant and fascinating is the development of the amazing analytical technology that has permitted us entry into the genomic era and a whole new level of scientific understanding. Analytical Techniques in DNA Sequencing takes a look at the various DNA
Author: David H. Persing Publisher: John Wiley & Sons ISBN: 1555819079 Category : Science Languages : en Pages : 835
Book Description
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.
Author: Erin Marie McElfish
Author: Erin Marie McElfish
Author: Keith R. Mitchelson
Author: 
Author: Ulrich Finckh
Author: Klaus D. Sattler
Author: Tom Strachan
Author: Shu Chien
Author: Brian K. Nunnally
Author: Robi David Mitra
Author: David H. Persing