Download or read book DNA Purification on Microfluidic Devices with a Focus on Large Volume Forensic Biological Samples written by Carmen Reneé Reedy and published by . This book was released on 2011 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Development of a Microfluidic System for Efficient DNA Purification from Large volume Blood Samples written by Jian Wen and published by . This book was released on 2007 with total page 336 pages. Available in PDF, EPUB and Kindle. Book excerpt: This dissertation strikes at the heart of one of the major challenges associated with developing microfluidic systems for genetic analysis---solid phase extraction (SPE) from real-world samples. The work described involves the development of a novel two-stage, dual-phase microfluidic device using a novel porous monolithic as the solid phase for high capacity DNA purification from human blood, arguably the most challenging of samples. The fundamental chemistry and microfluidic embodiment of this porous monolith is detailed, and the capacity of the monolith in the microfluidic-based device is determined as it pertains to whole blood DNA. To circumvent the competition between proteins and DNA in the blood sample for binding sites on the phase, a reversed-phase C18 pre-column is developed to selectively isolate the majority of the proteins from DNA present in the blood lysate upstream of the DNA capture phase. The enlisting of a protein capture phase as stage 1 coupled with the DNA extraction monolith in stage 2, the novel two-stage, dual-phase microfluidic device is involved. To meet the needs of clinical diagnostic community for large capacity DNA purification while minimizing the overall cost of device, a novel monolith surface passivation method is demonstrated. Finally, microchip design, specifically, the microchannel configuration and cross-sectional geometry, is optimized to efficiently eliminate DNA elution peak tailing, enabling for the faster release of DNA while still retaining the DNA concentrating effect of the SPE. Future directions are also suggested at the end of this dissertation.

Download or read book DNA Purification in Microfluidic Systems for Clinical and Forensic Application written by Joan Marie Bienvenue and published by . This book was released on 2007 with total page 416 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Closed System DNA Purification for Degraded Compromised Evidence in Microfluidic Devices written by James P. Landers and published by . This book was released on 2013 with total page 52 pages. Available in PDF, EPUB and Kindle. Book excerpt: "The aims of this grant were to develop a volume reduction solid phase extraction (vrSPE) microchip capable of accommodating large volumes. The device was then to be applied to degraded and compromised samples as well as the purification of mitochondrial DNA. The device was also to be demonstrated in a multiplexed format for as few as two samples. Finally devices were to be field tested in a forensic laboratory using non-probative evidentiary samples." -- Executive summary.

Download or read book DNA Extraction by Isotachophoresis in a Microfluidic Channel written by and published by . This book was released on 2011 with total page 12 pages. Available in PDF, EPUB and Kindle. Book excerpt: Biological assays have many applications. For example, forensics personnel and medical professionals use these tests to diagnose diseases and track their progression or identify pathogens and the host response to them. One limitation of these tests, however, is that most of them target only one piece of the sample - such as bacterial DNA - and other components (e.g. host genomic DNA) get in the way, even though they may be useful for different tests. To address this problem, it would be useful to extract several different substances from a complex biological sample - such as blood - in an inexpensive and efficient manner. This summer, I worked with Maxim Shusteff at Lawrence Livermore National Lab on the Rapid Automated Sample Prep project. The goal of the project is to solve the aforementioned problem by creating a system that uses a series of different extraction methods to extract cells, bacteria, and DNA from a complex biological sample. Biological assays can then be run on purified output samples. In this device, an operator could input a complex sample such as blood or saliva, and would receive separate outputs of cells, bacteria, viruses, and DNA. I had the opportunity to work this summer with isotachophoresis (ITP), a technique that can be used to extract nucleic acids from a sample. This technique is intended to be the last stage of the purification device. Isotachophoresis separates particles based on different electrophoretic mobilities. This technique is convenient for out application because free solution DNA mobility is approximately equal for DNA longer than 300 base pairs in length. The sample of interest - in our case DNA - is fed into the chip with streams of leading electrolyte (LE) and trailing electrolyte (TE). When an electric field is applied, the species migrate based on their electrophoretic mobilities. Because the ions in the leading electrolyte have a high electrophoretic mobility, they race ahead of the slower sample and trailing electrolyte ions. Conversely, the trailing electrolyte ions have a slow electrophoretic mobility, so they lag behind the sample, thus trapping the species of interest between the LE and TE streams. In a typical isotachophoresis configuration, the electric field is applied in a direction parallel to the direction of flow. The species then form bands that stretch across the width of the channel. A major limitation of that approach is that only a finite amount of sample can be processed at once, and the sample must be processed in batches. For our purposes, a form of free-flow isotachophoresis is more convenient, where the DNA forms a band parallel to the edges of the channel. To achieve this, in our chip, the electric field is applied transversely. This creates a force perpendicular to the direction of flow, which causes the different ions to migrate across the flow direction. Because the mobility of the DNA is between the mobility of the leading and the trailing electrolyte, the DNA is focused in a tight band near the center of the channel. The stream of DNA can then be directed to a different output to produce a highly concentrated outlet stream without batch processing. One hurdle that must be overcome for successful ITP is isolating the electrochemical reactions that result from the application of high voltage for the actual process of isotachophoresis. The electrochemical reactions that occur around metal electrodes produce bubbles and pH changes that are detrimental to successful ITP. The design of the chips we use incorporates polyacrylamide gels to serve as electrodes along the central channel. For our design, the metal electrodes are located away from the chip, and high conductivity buffer streams carry the potential to the chip, functioning as a 'liquid electrode.' The stream then runs alongside a gel barrier. The gel electrode permits ion transfer while simultaneously isolating the separation chamber from any contaminants in the outer, 'liquid electrode' streams. The difference in potential from one side of the chip to the other creates an electric field. This field traverses the inner, separation channel, containing the leading electrolyte, the trailing electrolyte, and the sample of interest (DNA). To increase the ease of use of the chips, a newer chip design has been fabricated. This design has wire electrodes integrated on the chip, rather than elsewhere. To keep the pH changes and bubbling isolated from the separation channel, the chip contains deeper wells near the electrodes so that the flowing buffer can wash away any gases that form around the electrode. This design is significantly more compact because it eliminates the cumbersome electrode boxes. Eliminating the electrode boxes also decreases the required voltage, making the experiments safer. This happens because when the 'liquid electrode' streams travel through small diameter tubing, they lose much of their voltage due to the electrical resistance of the fluid in the tubing.

Download or read book Tietz Textbook of Clinical Chemistry and Molecular Diagnostics E Book written by Nader Rifai and published by Elsevier Health Sciences. This book was released on 2017-01-16 with total page 4785 pages. Available in PDF, EPUB and Kindle. Book excerpt: The Tietz Textbook of Clinical Chemistry and Molecular Diagnostics, 6th Edition provides the most current and authoritative guidance on selecting, performing, and evaluating the results of new and established laboratory tests. This classic clinical chemistry reference offers encyclopedic coverage detailing everything you need to know, including: analytical criteria for the medical usefulness of laboratory tests, variables that affect tests and results, laboratory medicine, applications of statistical methods, and most importantly clinical utility and interpretation of laboratory tests. It is THE definitive reference in clinical chemistry and molecular diagnostics, now fully searchable and with quarterly content updates, podcasts, clinical cases, animations, and extended content online through Expert Consult. - Analytical criteria focus on the medical usefulness of laboratory procedures. - Reference ranges show new approaches for establishing these ranges — and provide the latest information on this topic. - Lab management and costs gives students and chemists the practical information they need to assess costs, allowing them to do their job more efficiently and effectively. - Statistical methods coverage provides you with information critical to the practice of clinical chemistry. - Internationally recognized chapter authors are considered among the best in their field. - Two-color design highlights important features, illustrations, and content to help you find information easier and faster. - NEW! Internationally recognized chapter authors are considered among the best in their field. - NEW! Expert Consult features fully searchable text, quarterly content updates, clinical case studies, animations, podcasts, atlases, biochemical calculations, multiple-choice questions, links to Medline, an image collection, and audio interviews. You will now enjoy an online version making utility of this book even greater. - UPDATED! Expanded Molecular Diagnostics section with 12 chapters that focus on emerging issues and techniques in the rapidly evolving and important field of molecular diagnostics and genetics ensures this text is on the cutting edge and of the most value. - NEW! Comprehensive list of Reference Intervals for children and adults with graphic displays developed using contemporary instrumentation. - NEW! Standard and international units of measure make this text appropriate for any user — anywhere in the world. - NEW! 22 new chapters that focus on applications of mass spectrometry, hematology, transfusion medicine, microbiology, biobanking, biomarker utility in the pharmaceutical industry and more! - NEW! Expert senior editors, Nader Rifai, Carl Wittwer and Rita Horvath, bring fresh perspectives and help ensure the most current information is presented. - UPDATED! Thoroughly revised and peer-reviewed chapters provide you with the most current information possible.

Download or read book Forensic DNA Analysis written by Catherine Cupples Connon and published by Springer Nature. This book was released on 2023-08-14 with total page 423 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume focuses on the latest techniques used in forensic DNA analysis. The chapters include a comprehensive collection of extraction, quantification, STR amplification, and detection methods for routine forensic samples, including manual, semi-automated, and automated procedures using both home-brew and commercial products. The chapters also discuss probabilistic modeling software and specialized start-to-finish procedures for mitochondrial DNA analysis, archived latent fingerprints, latent DNA, rapid DNA profiling, and next-generation sequencing. Written in the highly successful Methods in Molecular Biology series format, chapters include introduction to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and practical, Forensic DNA Analysis: Methods and Protocols is a valuable resource for researchers interested in learning more about forensic DNA analysis procedures.

Download or read book Tietz Textbook of Laboratory Medicine E Book written by Nader Rifai and published by Elsevier Health Sciences. This book was released on 2022-02-03 with total page 4232 pages. Available in PDF, EPUB and Kindle. Book excerpt: Use THE definitive reference for laboratory medicine and clinical pathology! Tietz Textbook of Laboratory Medicine, 7th Edition provides the guidance necessary to select, perform, and evaluate the results of new and established laboratory tests. Comprehensive coverage includes the latest advances in topics such as clinical chemistry, genetic metabolic disorders, molecular diagnostics, hematology and coagulation, clinical microbiology, transfusion medicine, and clinical immunology. From a team of expert contributors led by Nader Rifai, this reference includes access to wide-ranging online resources on Expert Consult — featuring the comprehensive product with fully searchable text, regular content updates, animations, podcasts, over 1300 clinical case studies, lecture series, and more. - Authoritative, current content helps you perform tests in a cost-effective, timely, and efficient manner; provides expertise in managing clinical laboratory needs; and shows how to be responsive to an ever-changing environment. - Current guidelines help you select, perform, and evaluate the results of new and established laboratory tests. - Expert, internationally recognized chapter authors present guidelines representing different practices and points of view. - Analytical criteria focus on the medical usefulness of laboratory procedures. - Use of standard and international units of measure makes this text appropriate for any user, anywhere in the world. - Elsevier eBooks+ provides the entire text as a fully searchable eBook, and includes animations, podcasts, more than 1300 clinical case studies, over 2500 multiple-choice questions, a lecture series, and more, all included with print purchase. - NEW! 19 additional chapters highlight various specialties throughout laboratory medicine. - NEW! Updated, peer-reviewed content provides the most current information possible. - NEW! The largest-ever compilation of clinical cases in laboratory medicine is included with print purchase on Elsevier eBooks+. - NEW! Over 100 adaptive learning courses included with print purchase on Elsevier eBooks+ offer the opportunity for personalized education.

Download or read book Issues in Industrial Applied and Environmental Chemistry 2011 Edition written by and published by ScholarlyEditions. This book was released on 2012-01-09 with total page 2599 pages. Available in PDF, EPUB and Kindle. Book excerpt: Issues in Industrial, Applied, and Environmental Chemistry: 2011 Edition is a ScholarlyEditions™ eBook that delivers timely, authoritative, and comprehensive information about Industrial, Applied, and Environmental Chemistry. The editors have built Issues in Industrial, Applied, and Environmental Chemistry: 2011 Edition on the vast information databases of ScholarlyNews.™ You can expect the information about Industrial, Applied, and Environmental Chemistry in this eBook to be deeper than what you can access anywhere else, as well as consistently reliable, authoritative, informed, and relevant. The content of Issues in Industrial, Applied, and Environmental Chemistry: 2011 Edition has been produced by the world’s leading scientists, engineers, analysts, research institutions, and companies. All of the content is from peer-reviewed sources, and all of it is written, assembled, and edited by the editors at ScholarlyEditions™ and available exclusively from us. You now have a source you can cite with authority, confidence, and credibility. More information is available at http://www.ScholarlyEditions.com/.

Download or read book Development of Microfluidic Modules for DNA Purification Via Phenol Extraction and Analyte Concentration Using Transverse Electrokinetics written by Mercedes C. Morales and published by . This book was released on 2012 with total page 122 pages. Available in PDF, EPUB and Kindle. Book excerpt: In this work, microfluidic platforms have been designed and evaluated to demonstrate microscale DNA purification via organic (phenol) extraction as well as analyte trapping and concentration using a transverse electrokinetic force balance. First, in order to evaluate DNA purification via phenol extraction in a microdevice, an aqueous phase containing protein and DNA and an immiscible receiving organic phase were utilized to evaluate microfluidic DNA extraction under both stratified and droplet-based flow conditions using a serpentine microfluidic device. The droplet based flow resulted in a significant improvement of protein partitioning from the aqueous phase due to the flow recirculation inside each droplet improving material convective transport into the organic phase. The plasmid recovery from bacterial lysates using droplet-based flow was high (>92%) and comparable to the recovery achieved using commercial DNA purification kits and standard macroscale phenol extraction. Second, a converging Y-inlet microfluidic channel with integrated coplanar electrodes was used to investigate transverse DNA and protein migration under uniform direct current (DC) electric fields. Negatively charged samples diluted in low and high ionic strength buffers were co-infused with a receiving buffer of the same ionic strength into a main channel where transverse electric fields were applied. Experimental results demonstrated that charged analytes could traverse the channel width and accumulate at the positive bias electrode in a low electroosmotic mobility and high electrophoretic mobility condition (high ionic strength buffer) or migrated towards an equilibrium position within the channel when both electroosmotic mobility and electrophoretic mobility are high (low ionic strength buffer). The different behaviors are the result of a balance between the electrophoretic force and a drag force induced by a recirculating electroosmotic flow generated across the channel width due to the bounding walls. The miniaturization of DNA phenol extraction and the novel electrokinetic trapping techniques presented in this research are the initial steps towards an efficient DNA sample preparation chip which could be integrated with post-extraction DNA manipulations for genomic analysis modules such as capillary electrophoretic separations.

Download or read book Forensic Biology written by Richard Li and published by CRC Press. This book was released on 2015-03-11 with total page 558 pages. Available in PDF, EPUB and Kindle. Book excerpt: Focusing on forensic serology and forensic DNA analysis, this book introduces students to the methods and techniques utilized by forensic biology laboratories. Using schematic illustrations to clarify concepts, this second edition explores the latest DNA profiling tools, contains three new chapters, and provides 200 new images. It also includes new tables for many chapters. Covering the full scope of forensic biology, the book uses an accessible style designed to enhance students education and training so they are prepared, both in the laboratory and in the field.

Download or read book Handbook of DNA Profiling written by Hirak Ranjan Dash and published by Springer. This book was released on 2022-06-29 with total page 1206 pages. Available in PDF, EPUB and Kindle. Book excerpt: This reference book comprehensively reviews the significance of DNA technology in forensic science. After presenting the theory, basic principles, tools and techniques that are used in forensic DNA typing, it summarizes various techniques, including autosomal STR, Y-STR, X-STR, mitochondrial DNA and NGS, used in solving both criminal as and civil cases, such as paternity disputes, identification of mutilated remains, and culprit identification in sexual assault and murder cases. It also provides an overview of DNA-based genetic diagnostics for various diseases, and discusses the role of DNA typing in drug reactions, as well as the application of non-human DNA profiling of animals and plants in forensic science investigations. Lastly, the book examines the role of internal quality control in maintaining the high quality of DNA profiling.

Download or read book Development of a Microfluidic Module for DNA Purification Via Phenol Extraction written by and published by . This book was released on 2008 with total page 65 pages. Available in PDF, EPUB and Kindle. Book excerpt: Purification of Deoxyribonucleic acid (DNA) by organic-aqueous liquid extraction, also called phenol extraction, is a standard technique commonly utilized in biology laboratories. In order to minimize interaction energies, membrane components and proteins naturally partition to the organic (phenol) phase while the DNA stays in the aqueous phase, where it can be easily removed. In recent years, microfluidics has become a driving force toward more efficient and autonomous platforms for fluid based diagnostics, chemical reaction chambers, separation and preparation of biological materials. In this work, the design, fabrication, and performance of long microfluidic devices for DNA extraction are presented. The devices were fabricated using soft lithography to transfer lithographically defined features into a PDMS structure via replica molding. Stratified-flow experiments using a rhodamine dye conjugated bovine serum albumin protein (BSA) in an aqueous phase were conducted to compare different microchannel designs based on their ability to remove proteins from the aqueous phase into the phenol phase. Additionally, the study of BSA partitioning and DNA isolation in a two-phase system under stratified flow condition were addressed, separately and conjunctly. Finally, protein partitioning and DNA recovery were analyzed to evaluate two types of mixing, passive diffusion through stratified flows and droplet enhancement mixing.

Download or read book Sample Extraction and Processing in Microfluidic DNA Analysis Devices written by Rongsheng Lin and published by . This book was released on 2005 with total page 280 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Molecular Forensics written by Ralph Rapley and published by John Wiley & Sons. This book was released on 2007-05-21 with total page 33 pages. Available in PDF, EPUB and Kindle. Book excerpt: Molecular Forensics offers a comprehensive coverage of the increasingly important role that molecular analysis plays within forensic science. Starting with a broad introduction of modern forensic molecular technologies, the text covers key issues from the initial scenes of crime sampling to the use of evidential material in the prosecution of legal cases. The book also explores the questions raised by the growing debate on the applications of national DNA databases and the resulting challenges of developing, maintaining and curating such vast data structures. The broader range of applications to non-human cases is also discussed, as are the statistical pitfalls of using so-called unique data such as DNA profiles, and the ethical considerations of national DNA databases. An invaluable reference for students taking courses within the Forensic and Biomedical sciences, and also useful for practitioners in the field looking for a broad overview of the subject. Provides a comprehensive overview of modern forensic molecular technologies. Explores the growing debate on the applications of national DNA databases. Discusses the initial phases of investigation to the conclusion of cases involving molecular forensic analysis.

Download or read book Microfluidic DNA Extraction and Purification from Forensic Samples written by Eugene Tan and published by . This book was released on 2015-02-16 with total page 92 pages. Available in PDF, EPUB and Kindle. Book excerpt: This work has been selected by scholars as being culturally important, and is part of the knowledge base of civilization as we know it. This work was reproduced from the original artifact, and remains as true to the original work as possible. Therefore, you will see the original copyright references, library stamps (as most of these works have been housed in our most important libraries around the world), and other notations in the work. This work is in the public domain in the United States of America, and possibly other nations. Within the United States, you may freely copy and distribute this work, as no entity (individual or corporate) has a copyright on the body of the work.As a reproduction of a historical artifact, this work may contain missing or blurred pages, poor pictures, errant marks, etc. Scholars believe, and we concur, that this work is important enough to be preserved, reproduced, and made generally available to the public. We appreciate your support of the preservation process, and thank you for being an important part of keeping this knowledge alive and relevant.

Download or read book Microfluidic Devices for Forensic DNA Analysis written by Jayna J. Shah and published by . This book was released on 2007 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: "The development of integrated, miniaturized, and portable DNA analysis systems is crucial to alleviate massive backlog of unanalyzed samples and to address ever increasing demand for these assays. This thesis work contributes towards the development of a fully integrated microdevice capable of 'sample in -- answer out' for forensic DNA analysis. Specifically, this work describes the development of rapid and robust fabrication protocol for solvent-actuated bonding of polymeric thermoplastic substrates at room temperature, the development of microchannel wall coating strategies to eliminate analyte-wall interactions for high resolution separation of single-stranded DNA, and the characterization of a thin-film planar microwave transmission line for microfluidic heating applications. The solvent-actuated bonding protocol was based on the difference in capillary forces between the microchannel and the interstitial space between the surfaces of the two substrates to be bonded. This force differential wicked the bonding solvent into the gap between the substrates causing them to bond. The technique was implemented by placing the two substrates under moderate pressure, applying a moderate pneumatic vacuum to the fluidic channel, and introducing tens of microliter of bonding solvent through one end of the fluidic channel. The effect of bonding solvent on the dimensions of the microchannel was analyzed, and the mechanical robustness of the bonded devices was also characterized. Electrophoretic separation of single-stranded DNA (ssDNA) was successfully performed to demonstrate the functionality of these devices. To enhance ssDNA separation performance, schemes to modify poly(methyl methacrylate) (PMMA) -- the primary substrate used in this work -- were explored. This two step process consisted of altering surface hydrophilicity via surface activation using either nitric acid or UV/ozone followed by coating the surfaces with adsorptive polymers. Contact-angle measurements of the pristine and modified PMMA substrates were performed to quantify the change in wettability of the surface. Twofold increase in the separation efficiency was achieved by implementing these surface passivation strategies. Finally, the use of a thin-film planar microwave transmission line as a microwave power source was investigated for on-chip microwave heating of fluids. The microwave characterization data was used to develop a first-order analytic model of the microwave power absorption. The model was used to understand microwave power flow through the device and to calculate the fraction of the incident power absorbed in the fluid. Additionally, a fit of the predicted temperature obtained using this model to the measured temperature was performed to evaluate efficiency of this heating method"--Abstract