Gene Synthesis By Assembly Of Short Oligonucleotides
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Gene Synthesis By Assembly Of Short Oligonucleotides
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Author :
language : en
Publisher:
Release Date : 2005
Gene Synthesis By Assembly Of Short Oligonucleotides written by and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2005 with categories.
In principle, a pre-constructed library of all possible short oligonucleotides could be used to construct many distinct gene sequences. This approach requires computational methods to accurately determine the assembly procedure, but relieves the current technological constraints of custom oligonucleotide synthesis. In order to assess the feasibility of such an approach, I examined T4 DNA Ligase activity on short oligonucleotides and found that ligation is dependent on the formation of a double-stranded DNA duplex of at least five base pairs flanking the site of ligation. However, ligations could be performed with overhangs smaller than five nucleotides and oligonucleotides as small as octamers, in the presence of a second, complementary oligonucleotide. As a proof of principle for DNA synthesis through the assembly of short oligonucleotides, I performed a hierarchical ligation procedure whereby octamers were combined to construct a target 128 bp segment of the human beta- ctin gene coding sequence. Thus, the construction of synthetic genes, without the need for custom oligonucleotide synthesis, is feasible. Algorithmic methods were then developed to extend this approach to DNA on the order of thousands of base pairs.
Towards An Inexpensive High Thoughroughput Method Of Automated Gene Synthesis
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Author : Michael John Bassetti
language : en
Publisher:
Release Date : 2007
Towards An Inexpensive High Thoughroughput Method Of Automated Gene Synthesis written by Michael John Bassetti and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2007 with categories.
Oligonucleotide Synthesis
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Author : Piet Herdewijn
language : en
Publisher: Springer Science & Business Media
Release Date : 2008-02-04
Oligonucleotide Synthesis written by Piet Herdewijn and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2008-02-04 with Medical categories.
A collection of powerful new techniques for oligonucleotide synthesis and for the use of modified oligonucleotides in biotechnology. Among the protocol highlights are a novel two-step process that yields a high purity, less costly, DNA, the synthesis of phosphorothioates using new sulfur transfer agents, the synthesis of LNA, peptide conjugation methods to improve cellular delivery and cell-specific targeting, and triple helix formation. The applications include using molecular beacons to monitor the PCR amplification process, nuclease footprinting to study the sequence-selective binding of small molecules of DNA, nucleic acid libraries, and the use of small interference RNA (siRNA) as an inhibitor of gene expression.
Gene Synthesis
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Author : Jean Peccoud
language : en
Publisher: Humana Press
Release Date : 2012-02-10
Gene Synthesis written by Jean Peccoud and has been published by Humana Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2012-02-10 with Medical categories.
The de novo fabrication of custom DNA molecules is a transformative technology that significantly affects the biotechnology industry. Basic genetic engineering techniques for manipulating DNA in vitro opened an incredible field of opportunity in the life sciences. In, Gene Synthesis: Methods and Protocols expert researchers in the field detail many of the methods which are now commonly used to fabricate DNA . These include methods and techniques for the assembly of oligonucleotide, cloning of synthons into larger fragments, protocols and software applications, and educational and biosecurity impacts of gene synthesis. Written in the highly successful Methods in Molecular BiologyTM series format, the chapters include the kind of detailed description and implementation advice that is crucial for getting optimal results in the laboratory. Thorough and intuitive, Gene Synthese: Methods and Protocols aids scientists in understanding all the different stages of a complex gene synthesis process, while refining their understanding of gene synthesis and determine what part of the process they can or should do in their laboratory and what parts should be contracted to a specialized service provider.
Synthesis Of Therapeutic Oligonucleotides
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Author : Satoshi Obika
language : en
Publisher: Springer
Release Date : 2018-12-08
Synthesis Of Therapeutic Oligonucleotides written by Satoshi Obika and has been published by Springer this book supported file pdf, txt, epub, kindle and other format this book has been release on 2018-12-08 with Medical categories.
This book presents the latest knowledge on a broad range of topics relating to the synthesis of natural and artificial oligonucleotides with therapeutic potential. Nucleic acid-based therapeutics are attracting much attention, and numerous therapeutic oligonucleotides, such as antisense oligonucleotides, siRNAs, splice-switching oligonucleotides, and nucleic acid aptamers, are being evaluated in clinical trials for the treatment of a variety of diseases. Synthesis of Therapeutic Oligonucleotides covers a broad range of topics in the field that are of high relevance to researchers, including the synthesis of natural and chemically modified oligonucleotides, the development of novel nucleic acid analogs, industrial scale synthesis and purification of oligonucleotides, and important aspects of chemistry, manufacturing, and controls (CMC). The aim is to provide new insights and inspire fresh ideas in nucleic acid chemistry that may ultimately lead to novel concepts and techniques and the discovery of more effective nucleic acid drugs. The book will be of high value for both established researchers in the field and students intending to specialize in nucleic acid chemistry research.
A Systematic And Extensible Approach To Dna Primer Design For Whole Gene Synthesis
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Author : Amanda Victrix Allen Wozniak
language : en
Publisher:
Release Date : 2005
A Systematic And Extensible Approach To Dna Primer Design For Whole Gene Synthesis written by Amanda Victrix Allen Wozniak and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2005 with categories.
The future of synthetic biology research hinges upon the development of accurate and inexpensive whole gene synthesis technologies. Recent advances in the purification of solid-phase manufactured oligonucleotides make it possible to manufacture whole genes by polymerase chain reaction methods. Yet, despite the improvement in laboratory methods, whole gene synthesis is not rapidly progressing because most gene design software takes an excessively naive approach to the complex problem of designing component oligonucleotides for whole gene synthesis. The synthetic biology community needs a flexible, robust and optimal primer design tool. We present the software design for a tool which designs oligonucleotides that are compatible with a wide variety of oligo purification and whole gene assembly protocols. Our design strategy uses physical sequence feature identification, optimal artificial intelligence search techniques, and sequence optimisation via intelligent codon substitution to produce near-optimal oligonucleotide arrays. We address all aspects of the oligonucleotide design problem, from physical constraints to the computational overhead involved in searching for an optimal solution, and provide an extensive set of data structures and algorithms.
Chemical And Enzymatic Synthesis Of Gene Fragments
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Author : Hans Günter Gassen
language : en
Publisher:
Release Date : 1982
Chemical And Enzymatic Synthesis Of Gene Fragments written by Hans Günter Gassen and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 1982 with Cell cycle categories.
Preface: In the past the chemical and enzymatic synthesis of oligonucleotides of defined sequence had to be left to a few experts. Now, however, with the triester approach, the phosphite method and the solid-support techniques gene fragment synthesis has turned into an easy procedure even for a non-chemist. Due to the elegant chemistry involved, all methods work without sophisticated equipment and are prone to mechanisation and eventual automation. It is hoped that combined chemical-encymatic gene synthesis may become a standard technique in a molecular biology laboratory, such as DNA sequencing or in-vitro recombination of nucleic acides. We omitted chemical RNA synthesis, since this field is developing so rapidly at the moment that one has to refer to the original publications. However, we included enzymatic synthesis of RNA fragments, procedures which already have obtained a high degree of standardisation. Most of the contributions are revised versions of the protocols supplied for the EMBO sponsored course on "Automated Chemical and Enzymic Gene Synthesis", held in Darmstadt, March 21 to April 3, 1982. The protocols were improved on the basis of the experience of 30 student scientists with chemical, biological or medical backgrounds. Previously omitted procedures, such as the wandering spot method for oligonucleotide analysis, were included. In editing the manuscript we encountered problems with the nomenclature of nucleic acid components. In unambiquous cases we favoured a simple description, hoping for example, that oligodeoxynucleotide is always understood to mean oligo-2'-deoxyribonucleotide. This book aims to provide those interested in DNA/RNA research with state-of-the-art methods in the synthesis, purification, and analysis of DNA and RNA fragments. The editors wish to thank the authors for their efforts in preparing manuscripts from the the revised laboratory protocols. We gratefully acknowledge the skill and the patience of Mrs. E. Ronnfeldt in typing the manuscripts. We express our thanks to Verlag Chemie for the friendly and very efficient cooperation.--H.G. Gassen A. Lang--Darmstadt, in July 1982.
Protocols For Oligonucleotides And Analogs
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Author : Sudhir Agrawal
language : en
Publisher: Springer Science & Business Media
Release Date : 1993-08-31
Protocols For Oligonucleotides And Analogs written by Sudhir Agrawal and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 1993-08-31 with Science categories.
When first conceived, not only was the aim of Protocols for Oligo nucleotides and Analogs to provide wide coverage of the ohgonuc- otide chemistry field for readers who are well versed within the field, but also to give investigators just entering into the field a new perspective. The very first book on this topic was edited and published by Michael Gait in 1984, in whose laboratory I encountered the newer aspects of oligonucleotide chemistry. Since then, oligonucleotide research has developed to such an extent that its uses extend far beyond basic studies, and now find wide application throughout clinical science as well. Until recently, the major application of oligonucleotides has been in the area of DNA-based diagnostic and "antisense oligonucleotid- based therapeutic approaches. However, oligonucleotides are now also being used as therapeutic agents and are thus frequently found in clinical trials in humans. Synthesis of unmodified oligonucleotides using automated synthe sizers has become a common practice in numerous laboratories. How ever, improvements on the existing techniques and the introduction of ever newer methods for oligonucleotide synthesis is constantly driving ahead in the leading research laboratories. And several new oligonucle otide analogs have been synthesized and studied for their individual prop erties in recent years. The present volume strives to bring the readers the most up-to-date information on the newest aspects of synthesis of oligo nucleotides and their analogs. A separate volume covers synthesis of oligonucleotide conjugates, along with most of the analytical techniques presently used for analysis of oligonucleotides.
Nucleic Acids In Chemistry And Biology
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Author : G Michael Blackburn
language : en
Publisher: Royal Society of Chemistry
Release Date : 2015-11-09
Nucleic Acids In Chemistry And Biology written by G Michael Blackburn and has been published by Royal Society of Chemistry this book supported file pdf, txt, epub, kindle and other format this book has been release on 2015-11-09 with Science categories.
The structure, function and reactions of nucleic acids are central to molecular biology and are crucial for the understanding of complex biological processes involved. Revised and updated Nucleic Acids in Chemistry and Biology 3rd Edition discusses in detail, both the chemistry and biology of nucleic acids and brings RNA into parity with DNA. Written by leading experts, with extensive teaching experience, this new edition provides some updated and expanded coverage of nucleic acid chemistry, reactions and interactions with proteins and drugs. A brief history of the discovery of nucleic acids is followed by a molecularly based introduction to the structure and biological roles of DNA and RNA. Key chapters are devoted to the chemical synthesis of nucleosides and nucleotides, oligonucleotides and their analogues and to analytical techniques applied to nucleic acids. The text is supported by an extensive list of references, making it a definitive reference source. This authoritative book presents topics in an integrated manner and readable style. It is ideal for graduate and undergraduates students of chemistry and biochemistry, as well as new researchers to the field.
Oligonucleotide Synthesis
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Author : Michael J. Gait
language : en
Publisher:
Release Date : 1984
Oligonucleotide Synthesis written by Michael J. Gait and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 1984 with Medical categories.