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 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 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 Analytical Advances in Forensic Science written by Katie Maree Horsman and published by . This book was released on 2007 with total page 372 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Forensic DNA Analysis written by Jaiprakash G. Shewale and published by CRC Press. This book was released on 2013-08-19 with total page 449 pages. Available in PDF, EPUB and Kindle. Book excerpt: The field of forensic DNA analysis has grown immensely in the past two decades and genotyping of biological samples is now routinely performed in human identification (HID) laboratories. Application areas include paternity testing, forensic casework, family lineage studies, identification of human remains, and DNA databasing. Forensic DNA Analysis: Current Practices and Emerging Technologies explores the fundamental principles and the application of technologies for each aspect of forensic DNA analysis. The book begins by discussing the value of DNA evidence and how to properly recognize, document, collect, and store it. The remaining chapters examine: The most widely adopted methods and the best practices for DNA isolation from forensic biological samples and human remains Studies carried out on the use of both messenger RNA and small (micro) RNA profiling Real-time polymerase chain reaction (PCR) methods for quantification and assessment of human DNA prior to genotyping Capillary electrophoresis (CE) as a tool for forensic DNA analysis Next-generation short tandem repeat (STR) genotyping kits for forensic applications, the biological nature of STR loci, and Y-chromosome STRs (Y-STRs) Mitochondrial DNA (mtDNA) sequence analysis Single nucleotide polymorphisms (SNPs) and insertion/deletion polymorphisms (indels) in typing highly degraded DNA Deep-sequencing technologies The current state of integrated systems in forensic DNA analysis The book concludes by discussing various aspects of sample-processing training and the entities that provide such training programs. This volume is an essential resource for students, researchers, teaching faculties, and other professionals interested in human identification/forensic DNA analysis.
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 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 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 Forensic DNA Typing Protocols written by Angel Carracedo and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 282 pages. Available in PDF, EPUB and Kindle. Book excerpt: A state-of-the-art collection of readily reproducible laboratory methods for DNA identity analysis, including Y chromosome haplotyping, mtDNA, and SNP typing. The book offers well-tested protocols for DNA quantification using real-time PCR on forensic samples and for the determination of the number of amelogenine gene copies. For forensic geneticists, there are readily reproducible methods for species identification, ancient DNA, and pharmacogenetics. Additional chapters address new applications in the forensic genetics lab, such a species identification or typing of CYP polymorphisms for the analysis of adverse to drugs.
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 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 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 Forensic DNA Analysis written by Jaiprakash G. Shewale and published by CRC Press. This book was released on 2013-08-19 with total page 445 pages. Available in PDF, EPUB and Kindle. Book excerpt: The field of forensic DNA analysis has grown immensely in the past two decades and genotyping of biological samples is now routinely performed in human identification (HID) laboratories. Application areas include paternity testing, forensic casework, family lineage studies, identification of human remains, and DNA databasing. Forensic DNA Analysis:
Download or read book Study of Microtip based Extraction and Purification of DNA from Human Samples for Portable Devices written by Gareth Fotouhi and published by . This book was released on 2015 with total page 147 pages. Available in PDF, EPUB and Kindle. Book excerpt: DNA sample preparation is essential for genetic analysis. However, rapid and easy-to-use methods are a major challenge to obtaining genetic information. Furthermore, DNA sample preparation technology must follow the growing need for point-of-care (POC) diagnostics. The current use of centrifuges, large robots, and laboratory-intensive protocols has to be minimized to meet the global challenge of limited access healthcare by bringing the lab to patients through POC devices. To address these challenges, a novel extraction method of genomic DNA from human samples is presented by using heat-cured polyethyleneimine-coated microtips generating a high electric field. The microtip extraction method is based on recent work using an electric field and capillary action integrated into an automated device. The main challenges to the method are: (1) to obtain a stable microtip surface for the controlled capture and release of DNA and (2) to improve the recovery of DNA from samples with a high concentration of inhibitors, such as human samples. The present study addresses these challenges by investigating the heat curing of polyethyleneimine (PEI) coated on the surface of the microtip. Heat-cured PEI-coated microtips are shown to control the capture and release of DNA. Protocols are developed for the extraction and purification of DNA from human samples. Heat-cured PEI-coated microtip methods of DNA sample preparation are used to extract genomic DNA from human samples. It is discovered through experiment that heat curing of a PEI layer on a gold-coated surface below 150°C could inhibit the signal of polymerase chain reaction (PCR). Below 150°C, the PEI layer is not completely cured and dissolved off the gold-coated surface. Dissolved PEI binds with DNA to inhibit PCR. Heat curing of a PEI layer above 150°C on a gold-coated surface prevents inhibition to PCR and gel electrophoresis. In comparison to gold-coated microtips, the 225°C-cured PEI-coated microtips improve the recovery of DNA to 45% efficiency. Furthermore, the 225°C-cured PEI-coated microtips recover more DNA than gold-coated microtips when the surface is washed. Heat-cured (225°C) PEI-coated microtips are used for the recovery of human genomic DNA from whole blood. A washing protocol is developed to remove inhibiting particles bound to the PEI-coated microtip surface after DNA extraction. From 1.25 [scientific symbol]L of whole blood, an average of 1.83 ng of human genomic DNA is captured, purified, and released using a 225°C-cured PEI-coated microtip in less than 30 minutes. The extracted DNA is profiled by short tandem repeat analysis (STR). For forensic and medical applications, genomic DNA is extracted from dried samples using heat-cured PEI-coated microtips that are integrated into an automated device. DNA extraction from dried samples is critical for forensics. The use of dried samples in the medical field is increasing because dried samples are convenient for storage, biosafety, and contamination. The main challenge is the time required to properly extract DNA in a purified form. Typically, a 1 hour incubation period is required to complete this process. Overnight incubation is sometimes necessary. To address this challenge, a pre-extraction washing step is investigated to remove inhibiting particles from dried blood spots (DBS) before DNA is released from dried form into solution for microtip extraction. The developed protocol is expanded to extract DNA from a variety of dried samples including nasal swabs, buccal swabs, and other forensic samples. In comparison to a commercial kit, the microtip-based extraction reduced the processing time from 1.5 hours to 30 minutes or less with an equivalent concentration of extracted DNA from dried blood spots. The developed assay will benefit genetic studies on newborn screening, forensic investigation, and POC diagnostics.
Download or read book Forensic DNA Profiling Protocols written by Patrick J. Lincoln and published by Springer Science & Business Media. This book was released on 1998-01-22 with total page 617 pages. Available in PDF, EPUB and Kindle. Book excerpt: This state-of-the-art collection of easily reproducible methods includes all of the major techniques of DNA analysis currently used in forensic identity testing. The methods include the recovery of DNA from a large range of sample types, analysis of DNA as single and multi-locus VNTR probes, PCR amplification of STR and other loci, and mitochondrial sequencing. The expert scientists writing here -- many from laboratories around the world -- also discuss how to interpret the results in cases of unknown identity and disputed parentage.-- Covers all steps from extraction of human DNA through to analysis and interpretation-- Takes advantage of new methodologies such as capillary electrophoresis-- Clear step-by-step instructions ensure unfailing reproducibility.
Download or read book Principles and Practices of DNA Analysis A Laboratory Manual for Forensic DNA Typing written by Hirak Ranjan Dash and published by Humana. This book was released on 2021-02-21 with total page 344 pages. Available in PDF, EPUB and Kindle. Book excerpt: The book presents hands-on protocols for conventional and advanced forensic DNA fingerprinting experiments. It includes manual, semi-automatic, and advanced automatic techniques for DNA extraction from different biological samples. It also discusses various qualitative and quantitative approaches for the assessment of extracted forensic DNA. It contains protocols for the amplification of short tandem repeat markers (STRs) for the amplification-based target enrichment of the forensic samples. Further, it examines genotyping of the STR loci through capillary electrophoresis and includes real-world case studies where forensic DNA analysis has been used in the criminal and civil disputes. The book concludes by presenting technological developments in the field of DNA forensic analysis. Suitable for beginners, it is a key reference resource on a wide variety of DNA profiling techniques and applications.
