Are you looking for read ebook online? Search for your book and save it on your Kindle device, PC, phones or tablets. Download Development Of A Pcr Free Direct Genomic Dna Detection Assay Using Rolling Circle Amplification PDF full book. Access full book title Development Of A Pcr Free Direct Genomic Dna Detection Assay Using Rolling Circle Amplification.
| Author | : Eric August Schopf |
| Publisher | : |
| Total Pages | : 298 |
| Release | : 2009 |
| Genre | : |
| ISBN | : |
| Author | : Vadim V. Demidov |
| Publisher | : Taylor & Francis |
| Total Pages | : 342 |
| Release | : 2004 |
| Genre | : Science |
| ISBN | : 9780954523299 |
Whereas most books on DNA amplification focus on PCR-based technologies, this volume presents a wider range of methods to amplify DNA with an emphasis on their diverse applications. The book covers both well-established and newly-developed protocols including ligation-based thermocycling approaches, real-time PCR and other new PCR developments, plus several powerful non-PCR isothermal DNA amplification techniques, for example: real-time strand displacement amplification (SDA), rolling-circle amplification (RCA) and multiple-displacement amplification (MDA). An entire section is devoted to a group of enzymes, both natural and engineered, which are employed for DNA amplification and related purposes. In addition, the use of DNA amplification in the detection of non-DNA analytes is presented.
| Author | : Giuseppe Spoto |
| Publisher | : Springer Science & Business Media |
| Total Pages | : 320 |
| Release | : 2012-07-06 |
| Genre | : Science |
| ISBN | : 940071226X |
This book offers an overview of state-of-the-art in non amplified DNA detection methods and provides chemists, biochemists, biotechnologists and material scientists with an introduction to these methods. In fact all these fields have dedicated resources to the problem of nucleic acid detection, each contributing with their own specific methods and concepts. This book will explain the basic principles of the different non amplified DNA detection methods available, highlighting their respective advantages and limitations. Non-amplified DNA detection can be achieved by adopting different techniques. Such techniques have allowed the commercialization of innovative platforms for DNA detection that are expected to break into the DNA diagnostics market. The enhanced sensitivity required for the detection of non amplified genomic DNA has prompted new strategies that can achieve ultrasensitivity by combining specific materials with specific detection tools. Advanced materials play multiple roles in ultrasensitive detection. Optical and electrochemical detection tools are among the most widely investigated to analyze non amplified nucleic acids. Biosensors based on piezoelectric crystal have been also used to detect unamplified genomic DNA. The main scientific topics related to DNA diagnostics are discussed by an outstanding set of authors with proven experience in this field.
| Author | : Simon Hughes |
| Publisher | : Scion Publishing Ltd |
| Total Pages | : 217 |
| Release | : 2005-09-01 |
| Genre | : Science |
| ISBN | : 1907904441 |
Whole genome amplification generates microgram quantities of genomic DNA starting from a sample of as little as a few femtograms and so is a vital technique when sample material is limited, as well as for high-throughput assays. Whole Genome Amplification: Methods Express is a comprehensive up-to-date laboratory manual for this key technique. It provides detailed step-by-step protocols as well as hints and tips for success and troubleshooting, taking readers through all aspects of whole genome amplification. This book is an essential practical guide for any researcher currently using PCR for genomic amplification or who wishes to do so in future.
| Author | : Yuvaneswari Chandramoulee Swaran |
| Publisher | : |
| Total Pages | : 0 |
| Release | : 2012 |
| Genre | : |
| ISBN | : |
The use of direct PCR with different types of sample was explored in this study. Genomic DNA preparations at various concentrations and buccal cell counts were deposited on commonly encountered substrates, recovered and amplified using direct PCR before subjecting them to capillary electrophoresis. The electropherograms obtained were compared to those obtained using the standard DNA profiling protocol which involves extraction and amplification prior to capillary electrophoresis. Direct PCR was found to be better than the standard DNA profiling protocol in both studies and was further tested with fingerprints, touch DNA on fabric and blood and semen stains on fabrics. All these tests were successful with direct PCR indicating that this technique has the potential to be incorporated into routine forensic DNA testing. Supplementary tests were also carried out to compare the efficiency of the swabbing technique utilised and the effect different substrates had on DNA recovery. Four non-porous substrates, which were glass, stainless steel, plastic and ceramic, and four types of dyed fabrics, which were white cotton, light blue denim, nylon and brown cotton, were used to deposit DNA and the resulting DNA profiles were evaluated. Of the non-porous substrates tested, the highest recovery of DNA was observed with plastic while the lowest was observed with stainless steel. DNA deposited on fabric on the other hand gave variable results which we believe is dependent on the dye used to stain the fabric and the thickness of the fibres used. The results in this experiment indicated that the substrate DNA is deposited on plays an important role in determining the resulting DNA profiles. Finally, a novel multiplex consisting of five autosomal and two Y-chromosomal STRs which also provides the inhibitor status of the sample was developed. This multiplex also addresses the issues concerning sensitivity and robustness that was encountered with other commercially available multiplexes. The multiplex was developed, validated and tested with various mock crime scene samples successfully. Allelic ladder, panels and bins were created to be used with this multiplex to aid in sample designation when subjected to capillary electrophoresis.
| Author | : Bimal D. M. Theophilus |
| Publisher | : Springer Science & Business Media |
| Total Pages | : 214 |
| Release | : 2008-02-02 |
| Genre | : Science |
| ISBN | : 1592592732 |
1Bimal D. Theophilus and Ralph Rapley provide biological and clinical investigators with a comprehensive collection of new, recent, and updated PCR-based screening methods suitable for detecting the presence of both known and novel mutations. The methods cover point mutations (e.g., ASO-PCR, SSCP, DGGE, chemical cleavage), deletions (multiplex PCR, FISH, blotting), non-sense mutations (PTT), and more. The new and exciting techniques of DNA array analysis, along with such recently developed experimental methods as conformation-sensitive gel electrophoresis, are also included. Each chapter explains the basic theory behind the technique and provides valuable notes essential for its successful execution.
| Author | : Marilyn J. Roossinck |
| Publisher | : Springer Science & Business Media |
| Total Pages | : 227 |
| Release | : 2008-02-23 |
| Genre | : Science |
| ISBN | : 3540757635 |
This book provides a comprehensive look at the field of plant virus evolution. It is the first book ever published on the topic. Individual chapters, written by experts in the field, cover plant virus ecology, emerging viruses, plant viruses that integrate into the host genome, population biology, evolutionary mechanisms and appropriate methods for analysis. It covers RNA viruses, DNA viruses, pararetroviruses and viroids, and presents a number of thought-provoking ideas.
| Author | : Kirstin J. Edwards |
| Publisher | : Taylor & Francis |
| Total Pages | : 362 |
| Release | : 2004 |
| Genre | : Polymerase chain reaction |
| ISBN | : 113418400X |
| Author | : Vadim V. Demidov |
| Publisher | : Springer |
| Total Pages | : 180 |
| Release | : 2016-11-09 |
| Genre | : Science |
| ISBN | : 331942226X |
This book covers the latest developments in rolling circle amplification (RCA) technology with applications in clinical diagnostic tests and molecular medicine. Topics covered include new enzymes useful in RCA, techniques involving RCA for enhanced signal amplification, novel RCA diagnostics, sensors for expediting RCA detection, and prospective RCA-based therapeutics. This is a valuable book for university professors and students in the field of biomedical engineering and biomolecular pharmacology as well as R&D managers of biotechnology and biopharmaceutical companies. Specifically, this book: Reviews prospective RCA-based therapeutics, including RCA-derived DNA nanoparticles that strongly bind to cancer cells Expands readers’ understanding of sensor systems for expediting detection of RCA products by using probe-tagged magnetic nanobeads Maximizes reader insights into novel RCA diagnostics, such as PNA openers-assisted RCA for detection of single target cells and in situ RCA diagnosis of cancer cells and malignant tissues Presents innovative methods for quasi-exponential enhancement of RCA-generated signals, such as nicking enzyme-assisted cascade RCA and RCA coupled with loop-mediated amplification Advance Praise for Rolling Circle Amplification (RCA): “This book provides a badly needed compendium of innovative RCA methods and applications. It should help further increase the community of scientists that have employed RCA in research and diagnostic programs.”— Charles Cantor, Professor Emeritus of Biomedical Engineering, Boston University Executive Director, Retrotope Inc. (USA) “In this new book Vadim Demidov has assembled an enticing menu of articles that illustrate the evolution of the RCA field, including improved protein parts for building superior DNA nanomachines, enhanced modalities of amplification and detection, diagnostic applications, and even a sampling of potential therapeutic applications. The reader will appreciate that while RCA has come of age, there is no lack of exciting surprises, turns, and twists in the continuing evolution of the technology.”— Paul Lizardi, Professor of Pathology, Yale University School of Medicine (retired) Investigator, University of Granada, Spain, President, PetaOmics, Inc., San Marcos, Texas.
| Author | : Erin Marie McElfish |
| Publisher | : |
| Total Pages | : 77 |
| Release | : 2009 |
| Genre | : |
| ISBN | : |
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.
