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 DNA Binding and DNA Extraction written by Chunxu Zhou and published by Nova Science Publishers. This book was released on 2012-03 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: In this book, the authors present current research in the study of DNA binding and extraction methods, applications, and limitations. Topics discussed in this compilation include DNA binding to cationic polymers and its implications on gene therapy; metal based anticancer drug-DNA binding; non-covalent DNA-AuNP's interactions; prospects of nanoparticle DNA binding; extraction and purification of DNA from USAB reactor sludge samples; isolation of genomic DNA from microbes and microbial communities; dengue virus RNA extraction challenges and isolating bacterial and viral nucleic acids from clinical samples for diagnostic testing.
Download or read book Microchip Capillary Electrophoresis written by Charles Henry and published by Humana. This book was released on 2006-03-01 with total page 238 pages. Available in PDF, EPUB and Kindle. Book excerpt: Leading chemists and engineers concisely explain the principles behind microchip capillary electrophoresis and demonstrate its use in a variety of biochemical applications, ranging from the analysis of DNA, proteins, and peptides to single cell analysis and measuring the impact of surface modification on flow in microfluidic channels. Since surface chemistry must be carefully considered for optimal operation at this scale, the authors also discuss methods of both adsorbed and covalent surface modification for its control. Fabrication methods for producing microchips with glass, poly(dimethylsiloxane), and other polymers are also provided so that even novices can produce simple devices for standard separations. Microchip Capillary Electrophoresis: Methods and Protocols provides a practical starting point for either initiating research in the field of microchip capillary electrophoresis or understanding the full range of what can be done with existing systems.
Download or read book Medical Biosensors for Point of Care POC Applications written by Roger Narayan and published by Woodhead Publishing. This book was released on 2016-08-21 with total page 318 pages. Available in PDF, EPUB and Kindle. Book excerpt: Medical Biosensors for Point of Care (POC) Applications discusses advances in this important and emerging field which has the potential to transform patient diagnosis and care. Part 1 covers the fundamentals of medical biosensors for point-of-care applications. Chapters in part 2 go on to look at materials and fabrication of medical biosensors while the next part looks at different technologies and operational techniques. The final set of chapters provide an overview of the current applications of this technology. Traditionally medical diagnostics have been dependent on sophisticated technologies which only trained professionals were able to operate. Recent research has focused on creating point-of-care diagnostic tools. These biosensors are miniaturised, portable, and are designed to be used at the point-of-care by untrained individuals, providing real-time and remote health monitoring. - Provides essential knowledge for designers and manufacturers of biosensors for point-of-care applications - Provides comprehensive coverage of the fundamentals, materials, technologies, and applications of medical biosensors for point-of-care applications - Includes contributions from leading international researchers with extensive experience in developing medical biosensors - Discusses advances in this important and emerging field which has the potential to transform patient diagnosis and care
Download or read book Basic Techniques in Molecular Biology written by Stefan Surzycki and published by Springer Science & Business Media. This book was released on 2012-12-06 with total page 442 pages. Available in PDF, EPUB and Kindle. Book excerpt: This laboratory manual gives a thorough introduction to basic techniques. It is the result of practical experience, with each protocol having been used extensively in undergraduate courses or tested in the authors laboratory. In addition to detailed protocols and practical notes, each technique includes an overview of its general importance, the time and expense involved in its application and a description of the theoretical mechanisms of each step. This enables users to design their own modifications or to adapt the method to different systems. Surzycki has been holding undergraduate courses and workshops for many years, during which time he has extensively modified and refined the techniques described here.
Download or read book Protein Analysis and Purification written by I.M. Rosenberg and published by Springer Science & Business Media. This book was released on 2013-03-14 with total page 453 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book is designed to be a practical progression of experimental techniques an investigator may follow when embarking on a biochemical project. The protocols may be performed in the order laid out or may be used inde pendently. The aim of the book is to assist a wide range of researchers. from the novice to the frustrated veteran, in the choice and design of experiments that are to be performed to provide answers to specific questions. The manual describes standard techniques that have been shown to work, as well as some newer ones that are beginning to prove important. By following the promi nently numbered steps. you can work your way through any protocol. whether it's a new technique or a task you've done before for which you need a quick review or updated methodology. This manual will assist the experimentalist in designing properly controlled experiments. There will be no advice for dealing with specific pieces of equip ment other than encouragement to read the manual, if you can find it. Through out all manipulations try to be objective. Be on the lookout for unexpected findings. You will learn the most from unexpected results. and they are often the beginning of the next project. It is never possible to record too much in your lab notebook. Do not get discouraged. Remember, things will not always run smoothly.
Download or read book Gene Biotechnology written by William Wu and published by CRC Press. This book was released on 2016-04-19 with total page 576 pages. Available in PDF, EPUB and Kindle. Book excerpt: Covering state-of-the-art technologies and a broad range of practical applications, the Third Edition of Gene Biotechnology presents tools that researchers and students need to understand and apply today's biotechnology techniques. Many of the currently available books in molecular biology contain only protocol recipes, failing to explain the princ
Download or read book Sample Preparation Techniques in Analytical Chemistry written by Somenath Mitra and published by John Wiley & Sons. This book was released on 2004-04-07 with total page 480 pages. Available in PDF, EPUB and Kindle. Book excerpt: The importance of accurate sample preparation techniques cannot be overstated--meticulous sample preparation is essential. Often overlooked, it is the midway point where the analytes from the sample matrix are transformed so they are suitable for analysis. Even the best analytical techniques cannot rectify problems generated by sloppy sample pretreatment. Devoted entirely to teaching and reinforcing these necessary pretreatment steps, Sample Preparation Techniques in Analytical Chemistry addresses diverse aspects of this important measurement step. These include: * State-of-the-art extraction techniques for organic and inorganic analytes * Sample preparation in biological measurements * Sample pretreatment in microscopy * Surface enhancement as a sample preparation tool in Raman and IR spectroscopy * Sample concentration and clean-up methods * Quality control steps Designed to serve as a text in an undergraduate or graduate level curriculum, Sample Preparation Techniques in Analytical Chemistry also provides an invaluable reference tool for analytical chemists in the chemical, biological, pharmaceutical, environmental, and materials sciences.
Download or read book Electrochemical Biosensors written by Serge Cosnier and published by CRC Press. This book was released on 2015-01-26 with total page 405 pages. Available in PDF, EPUB and Kindle. Book excerpt: Since four decades, rapid detection and monitoring in clinical and food diagnostics and in environmental and biodefense have paved the way for the elaboration of electrochemical biosensors. Thanks to their adaptability, ease of use in relatively complex samples, and their portability, electrochemical biosensors now are one of the mainstays of analy
Download or read book Proteomic Profiling and Analytical Chemistry written by Pawel Ciborowski and published by Elsevier. This book was released on 2016-03-02 with total page 300 pages. Available in PDF, EPUB and Kindle. Book excerpt: Proteomic Profiling and Analytical Chemistry: The Crossroads, Second Edition helps scientists without a strong background in analytical chemistry to understand principles of the multistep proteomic experiment necessary for its successful completion. It also helps researchers who do have an analytical chemistry background to break into the proteomics field. Highlighting points of junction between proteomics and analytical chemistry, this resource links experimental design with analytical measurements, data analysis, and quality control. This targeted point of view will help both biologists and chemists to better understand all components of a complex proteomic study. The book provides detailed coverage of experimental aspects such as sample preparation, protein extraction and precipitation, gel electrophoresis, microarrays, dynamics of fluorescent dyes, and more. The key feature of this book is a direct link between multistep proteomic strategy and quality control routinely applied in analytical chemistry. This second edition features a new chapter on SWATH-MS, substantial updates to all chapters, including proteomic database search and analytical quantification, expanded discussion of post-hoc statistical tests, and additional content on validation in proteomics. - Covers the analytical consequences of protein and peptide modifications that may have a profound effect on how and what researchers actually measure - Includes practical examples illustrating the importance of problems in quantitation and validation of biomarkers - Helps in designing and executing proteomic experiments with sound analytics
Download or read book Peptidomics written by Michael Schrader and published by Humana. This book was released on 2019-06-04 with total page 423 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume describes protocols for basic state-of-the-art approaches in the field of peptidomics. Most of these approaches are independent of the instruments used for analysis and can easily be adapted for equipment that is available in a typical proteomics facility. Chapters detail many of the basic techniques used to detect and identify peptides, methods for the relative quantitation of peptides between samples using isotopic labels or label-free approaches, and biological species as well as sample types. Written in the highly successful format of the Methods in Molecular Biology series, each chapter includes an introduction to the topic, a list of the necessary materials and reagents, reproducible step-by-step laboratory protocols, and tips on troubleshooting common problems and avoiding pitfalls. Authoritative and practical, Peptidomics: Methods and Strategies provides useful guidance for studies in the rapidly growing field of peptidomics.
Download or read book Plant Molecular Biology Manual written by Stanton Gelvin and published by Springer Science & Business Media. This book was released on 2013-11-11 with total page 138 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Manual of Minor Oral Surgery for the General Dentist written by Pushkar Mehra and published by John Wiley & Sons. This book was released on 2015-08-03 with total page 317 pages. Available in PDF, EPUB and Kindle. Book excerpt: The Manual of Minor Oral Surgery for the General Dentist, Second Edition continues the aim of providing clear and practical guidance to common surgical procedures encountered in general practice. Fully revised and updated with three additional chapters, the book approaches each procedure through detailed, step-by-step description and illustration. Ideal for general dental practitioners and students, the book is an indispensible tool for planning, performing, and evaluating a range of surgical procedures in day-to-day practice. The Manual of Minor Oral Surgery for the General Dentist begins with an expanded chapter on patient evaluation and history taking and a new chapter on managing the patient with medical comorbidities. It also address infections and sedation besides procedural chapters on such topics as third molar extractions, preprosthetic surgery, surgical implantology, crown-lengthening, and biopsy of oral lesions.
Download or read book Environmental Chemical Analysis written by S. Mitra and published by CRC Press. This book was released on 2018-10-03 with total page 345 pages. Available in PDF, EPUB and Kindle. Book excerpt: The study of the environment requires the reliable and accurate measurement of extremely small quantities of chemicals and the ability to determine if they are pollutants or naturally occurring species. Historically, a "dilute and disperse" method of waste disposal has been accepted; yet as we learn the long-term consequences of such an approach, it is clear that more rigorous waste management techniques are necessary to understand the sources and fates of contaminants and to regulate their discharge. This volume presents the details of the basic analytical science involved in making these measurements. It concentrates on the basic principles of sampling and sample preparation, followed by the chemical principles of the major instrumental methods used in chemical analysis, and detailed discussions of the major environmental matrices. This book also provides coverage of topics usually only partially discussed in textbooks, such as quality assurance plans and statistical data handling. Students majoring in environmental sciences need a foundation in measurement techniques used in the field. Environmental Chemical Analysis gives students a thorough grounding in this field and enough information to judge the quality and interpret the information produced in the analytical laboratory.
Download or read book Modern Proteomics Sample Preparation Analysis and Practical Applications written by Hamid Mirzaei and published by Springer. This book was released on 2016-12-14 with total page 525 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume serves as a proteomics reference manual, describing experimental design and execution. The book also shows a large number of examples as to what can be achieved using proteomics techniques. As a relatively young area of scientific research, the breadth and depth of the current state of the art in proteomics might not be obvious to all potential users. There are various books and review articles that cover certain aspects of proteomics but they often lack technical details. Subject specific literature also lacks the broad overviews that are needed to design an experiment in which all steps are compatible and coherent. The objective of this book was to create a proteomics manual to provide scientists who are not experts in the field with an overview of: 1. The types of samples can be analyzed by mass spectrometry for proteomics analysis. 2. Ways to convert biological or ecological samples to analytes ready for mass spectral analysis. 3. Ways to reduce the complexity of the proteome to achieve better coverage of the constituent proteins. 4. How various mass spectrometers work and different ways they can be used for proteomics analysis 5. The various platforms that are available for proteomics data analysis 6. The various applications of proteomics technologies in biological and medical sciences This book should appeal to anyone with an interest in proteomics technologies, proteomics related bioinformatics and proteomics data generation and interpretation. With the broad setup and chapters written by experts in the field, there is information that is valuable for students as well as for researchers who are looking for a hands on introduction into the strengths, weaknesses and opportunities of proteomics.
Download or read book Biochemical Plant Pathology written by J. A. Callow and published by . This book was released on 1983-01-01 with total page 496 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Pharmacogenomics written by Federico Innocenti and published by Humana Press. This book was released on 2013-07-04 with total page 359 pages. Available in PDF, EPUB and Kindle. Book excerpt: Understanding an individual's genetic makeup is the key to creating personalized drugs with greater efficacy and safety, and pharmacogenomics aims to study the complex genetic basis of inter-patient variability in response to drug therapy. Based upon the success of its first edition, the second edition of Pharmacogenomics: Methods And Protocols aims to continue providing readers with high-quality content on the most innovative and commonly adopted technologies in the field of pharmacogenomics as presented by experts in the field. Broken into several sections, this detailed volume examines techniques for interrogating variation in human genes and genomes, functional assessment of genetic variation, both in vitro and in vivo, as well as tools for translation and implementation of pharmacogenetic markers. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to the respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Comprehensive and thoroughly updated, Pharmacogenomics: Methods And Protocols, Second Edition serves as an essential reference and an invaluable source on the latest information in this field.
