Download or read book Method of Preparing and Applying Single Stranded DNA Probes to Double Stranded Target DNAs in Situ written by and published by . This book was released on 1991 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: A method is provided for producing single stranded non-self-complementary nucleic acid probes, and for treating target DNA for use therewith. Probe is constructed by treating DNA with a restriction enzyme and an exonuclease to form template/primers for a DNA polymerase. The digested strand is resynthesized in the presence of labeled nucleoside triphosphate precursor. Labeled single stranded fragments are separated from the resynthesized fragments to form the probe. Target DNA is treated with the same restriction enzyme used to construct the probe, and is treated with an exonuclease before application of the probe. The method significantly increases the efficiency and specificity of hybridization mixtures by increasing effective probe concentration by eliminating self-hybridization between both probe and target DNAs, and by reducing the amount of target DNA available for mismatched hybridizations.
Download or read book Official Gazette of the United States Patent and Trademark Office written by United States. Patent and Trademark Office and published by . This book was released on 2001 with total page 1312 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Nonisotopic DNA Probe Techniques written by Larry J. Kricka and published by Academic Press. This book was released on 2012-12-02 with total page 373 pages. Available in PDF, EPUB and Kindle. Book excerpt: Recently many nonisotopic methods of probing specific DNA sequences have been developed as replacements for radioactive labels, such as 32phosphorous and 125iodine. This book brings all of these new methods together in one convenient, easily accessible source. It enables researchers to select the nonisotopic method best suited to their application and to use it to maximum advantage by following the straightforward instructions provided. This book contains chapters on colorimetric, bioluminescent, chemiluminescent, fluorescent, and time-resolved fluorescent detection methods. Each chapter has been written by the inventor or developer of a particular nonisotopic method and thus provides an expert account of the method. Each chapterpresents useful background information and detailed, step-by-step, easy-to-follow, experimental procedures for labeling and detection. Gives extensive practical information Covers major types of nonisotopic labels and procedures Presents background information for each method Provides strategies and detailed experimental procedures for labeling and detecting DNA sequences by Fluorescence Chemiluminescence Bioluminescence Colorimetry
Download or read book Method for Producing Labeled Single stranded Nucleic Acid Probes written by and published by . This book was released on 1999 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: Disclosed is a method for the introduction of unidirectional deletions in a cloned DNA segment. More specifically, the method comprises providing a recombinant DNA construct comprising a DNA segment of interest inserted in a cloning vector, the cloning vector having an f1 endonuclease recognition sequence adjacent to the insertion site of the DNA segment of interest. The recombinant DNA construct is then contacted with the protein pII encoded by gene II of phage f1 thereby generating a single-stranded nick. The nicked DNA is then contacted with E. coli Exonuclease III thereby expanding the single-stranded nick into a single-stranded gap. The single-stranded gapped DNA is then contacted with a single-strand-specific endonuclease thereby producing a linearized DNA molecule containing a double-stranded deletion corresponding in size to the single-stranded gap. The DNA treated in this manner is then incubated with DNA ligase under conditions appropriate for ligation. Also disclosed is a method for producing single-stranded DNA probes. In this embodiment, single-stranded gapped DNA, produced as described above, is contacted with a DNA polymerase in the presence of labeled nucleotides to fill in the gap. This DNA is then linearized by digestion with a restriction enzyme which cuts outside the DNA segment of interest. The product of this digestion is then denatured to produce a labeled single-stranded nucleic acid probe.
Download or read book DNA Probes written by George H. Keller and published by . This book was released on 1993 with total page 696 pages. Available in PDF, EPUB and Kindle. Book excerpt: This edition is an updated version of the first edition, plus many new sections on specific applications of nucleic acid probes. Due to the explosion of such applications, experts in each major area have described their specialty. As in the first edition, the emphasis is on the commercial uses of DNA probes, as in diagnostic applications, and it incorporates background material, advice and specific protocols.
Download or read book DNA Probes written by George H. Keller and published by . This book was released on 1989 with total page 288 pages. Available in PDF, EPUB and Kindle. Book excerpt: This text concentrates on DNA probes, providing background information, sample preparation, isotopic labeling procedures, non-isotopic labeling procedures, hybridization formats and detection procedures, and amplification systems.
Download or read book Employing Double stranded DNA Probes on Colloidal Substrates for Competitive Hybridization Events written by Bryan Alexander Baker and published by . This book was released on 2010 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: The study of the DNA has found application beyond our understanding of its cellular function and into a variety of materials assembly and nucleic acid detection systems. The current research investigates double-stranded DNA probes in both a colloidal particle assembly and fluorescent assay format utilizing competitive hybridization events. In both contexts, the affinity of the dsProbes is tuned by the sequence design parameters of duplex length and complementarity. These systems were incubated with nucleic acid targets of interest and, based on the mechanism of competitive hybridization, were responsive to the presence of a high affinity competitive target. In the case of the particle assemblies, incubation with the competitive target resulted in observable disassembly of particle structures. In the case of fluorescently labeled dsProbes, incubation with competitive targets resulted in a quantifiable loss of fluorescence as determined by flow cytometry. Utilizing the fluorescently labeled dsProbe system, the kinetics of competitive hybridization was characterized for nucleic acid targets of varying specificity and strand context. The results indicate promise for the development of the competitive hybridization approach in nucleic acid detection systems providing advantages over current single-stranded probe designs. By utilizing a fluorescently labeled dsProbe approach, it is unnecessary to chemically modify the target of interest to impart a signaling mechanism. Additionally, as the process of competitive hybridization of dsProbes with targets of interest is an affinity driven process, discrimination of targets based on specificity is decoupled from standard measures such as elevated temperature protocols, an important step in translating nucleic acid technologies from the controlled laboratory environment to field applications.
Download or read book Medical BioMethods Handbook written by John M. Walker and published by Springer Science & Business Media. This book was released on 2005-03-18 with total page 635 pages. Available in PDF, EPUB and Kindle. Book excerpt: John Walker and Ralph Rapley have collected a wide-ranging group of molecular and biochemical techniques that are the most frequently used in medical and clinical research, especially diagnostics. The authors-well-established investigators who run their own research programs and use the methods on a regular basis-outline the practical procedures for using them and describe a variety of pertinent applications. Among the technologies presented are southern and western blotting, electrophoresis, PCR, cDNA and protein microarrays, liquid chromatography, in situ hybridization, karyotyping, flow cytometry, bioinformatics, genomics, and ribotyping. The applications include assays for mutation detection, mRNA analysis, chromosome translocations, inborn errors of metabolism, protein therapeutics, and gene therapy.
Download or read book Multiplex Target Capture with Double Stranded DNA Probes written by dna and published by . This book was released on 2016-01-15 with total page 36 pages. Available in PDF, EPUB and Kindle. Book excerpt: Target enrichment technologies utilize single-stranded oligonucleotide probes to capture candidate genomic regions from a DNA sample before sequencing. We describe target capture using double-stranded probes, which consist of single-stranded, complementary long padlock probes (cLPPs), each selectively capturing one strand of a genomic target through circularization. Using two probes per target increases sensitivity for variant detection and cLPPs are easily produced by PCR at low cost. Additionally, we introduce an approach for generating capture libraries with uniformly randomized template orientations. This facilitates bidirectional sequencing of both the sense and antisense template strands during one paired-end read, which maximizes target coverage.
Download or read book Exploiting the Energy of Intercalation for Sequence specific Recognition of Double stranded DNA written by Dale C. Guenther and published by . This book was released on 2015 with total page 454 pages. Available in PDF, EPUB and Kindle. Book excerpt: There has been great interest in developing probes capable of recognizing specific regions of double-stranded DNA (dsDNA) due to the vast potential of applications in the fields of molecular biology, genomic diagnostics, and DNA nanotechnology. Early efforts focused on targeting the accessible grooves of the DNA helix by means of triplex forming oligonucleotides (TFOs) or pyrrole/imidazole polyamides. However, such attempts have been limited by sequence restrictions and/or requirement for non-physiological conditions. A conceptually more elegant, but also more challenging, approach is to target the buried base pairs of the duplex and utilize the intrinsic Watson-Crick binding rules of nucleic acids. Single-stranded peptide nucleic acids (PNA) have a neutral backbone and bind to complementary DNA with very high affinity, facilitating duplex invasion, albeit at low ionic strengths. We have introduced Invader probes as an alternative approach for recognition of dsDNA. A mini-review of this approach is presented in CHAPTER 1. These probes are modified double-stranded oligonucleotides that are energetically activated for dsDNA recognition due to the low stability of the probe duplex and high affinity duplexes of each individual probe strand towards complementary dsDNA. The driving force for dsDNA recognition is the result of 2'-intercalator-functionalized nucleotides positioned in +1 interstrand zipper arrangements, termed 'energetic hotspots'. Major efforts in the laboratory have been focused on optimizing the chemistry of the monomers, while my projects primarily focused on the optimization of the probe architecture and applications of Invader probes. Initially, I set out to determine the influence of the number, location, and distance between the energetic hotspots. Optimal probe designs were then used for sequence-specific recognition of chromosomal DNA in a non-denaturing fluorescence in situ hybridization assay (CHAPTER 2). I then introduce bulged Invader probes, which feature an alkyl bulge in the double-stranded probe, which induces fraying and localized perturbation, resulting in faster and more efficient recognition of dsDNA (CHAPTER 3 and 4). Invader probes capable of photoactivated interstrand cross-linking are presented in APPENDIX A, which results in a covalent bond between the probe and DNA strands, resulting in more persistent binding. Finally, I evaluated Invader probes for inhibition of in vitro transcription (APPENDIX B).
Download or read book Genetic Engineering Principles and Methods written by Jane Setlow and published by Springer Science & Business Media. This book was released on 2012-12-06 with total page 361 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book FISH Technology written by Bernd W. Rautenstrauß and published by Springer Science & Business Media. This book was released on 2012-12-06 with total page 493 pages. Available in PDF, EPUB and Kindle. Book excerpt: Fluorescence in situ hybridization (FISH) has been developed as a powerful technology which allows direct visualisation or localisation of genomic alterations. The technique has been adopted to a range of applications in both medicine, especially in the areas of diagnostic cytogenetics, and biology. Topics described in this manual include: FISH on native human tissues, such as blood, bone marrow, epithelial cells, hair root cells, amniotic fluid cells, human sperm cells; FISH on archival human tissues, such as formalin fixed and paraffin embedded tissue sections, cryofixed tissue; simultaneous detection of apoptosis and xpression of apoptosis-related genes; comparative genomic ybridization; and special FISH techniques.
Download or read book The Development and Application of a Sequence selective DNA Sensor written by Kelly M. Millan and published by . This book was released on 1997 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book In Situ Hybridization Methods written by Giselbert Hauptmann and published by Humana. This book was released on 2016-10-05 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume contains a comprehensive compilation of chromogenic and fluorescent RNA in situ hybridization (ISH) technology in many of its various shades, forms, and applications. The book is organized into a number of parts and chapters focusing on the application of ISH methodologies to different animal species as used in Evolutionary Development (EvoDevo) and Biomedical research, and covering new developments in RNA visualization by fluorescent ISH (FISH). The described (F)ISH protocols employ effective strategies for signal enhancement and target amplification allowing for high signal intensities and drastically improved signal-to-noise ratios. Chromogenic and fluorescent ISH, as specified in the various chapters, are most essential for RNA expression profiling, applied to many fields of research including cellular, developmental, and evolutionary biology, neurobiology and neuropathology. Written for the popular Neuromethods series, chapters include the kind of detail and key implementation advice that ensures successful results in the laboratory. Essential and authoritative, In Situ Hybridization Methods provides detailed protocols for newcomers to ISH, and inspires researchers familiar with the technique to seek and find up-to-date methodology for new and specialized applications.
Download or read book Human Biochemistry written by Gerald Litwack and published by Academic Press. This book was released on 2021-11-28 with total page 883 pages. Available in PDF, EPUB and Kindle. Book excerpt: Human Biochemistry, Second Edition provides a comprehensive, pragmatic introduction to biochemistry as it relates to human development and disease. Here, Gerald Litwack, award-wining researcher and longtime teacher, discusses the biochemical aspects of organ systems and tissue, cells, proteins, enzymes, insulins and sugars, lipids, nucleic acids, amino acids, polypeptides, steroids, and vitamins and nutrition, among other topics. Fully updated to address recent advances, the new edition features fresh discussions on hypothalamic releasing hormones, DNA editing with CRISPR, new functions of cellular prions, plant-based diet and nutrition, and much more. Grounded in problem-driven learning, this new edition features clinical case studies, applications, chapter summaries, and review-based questions that translate basic biochemistry into clinical practice, thus empowering active clinicians, students and researchers. Presents an update on a past edition winner of the 2018 Most Promising New Textbook (College) Award (Texty) from the Textbook and Academic Authors Association and the PROSE Award of the Association of American Publishers Provides a fully updated resource on current research in human and medical biochemistry Includes clinical case studies, applications, chapter summaries and review-based questions Adopts a practice-based approach, reflecting the needs of both researchers and clinically oriented readers
Download or read book Index of Patents Issued from the United States Patent and Trademark Office written by and published by . This book was released on 1991 with total page 1896 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Encyclopedia of Food Microbiology written by Carl A. Batt and published by Academic Press. This book was released on 2014-04-02 with total page 3243 pages. Available in PDF, EPUB and Kindle. Book excerpt: Written by the world's leading scientists and spanning over 400 articles in three volumes, the Encyclopedia of Food Microbiology, Second Edition is a complete, highly structured guide to current knowledge in the field. Fully revised and updated, this encyclopedia reflects the key advances in the field since the first edition was published in 1999 The articles in this key work, heavily illustrated and fully revised since the first edition in 1999, highlight advances in areas such as genomics and food safety to bring users up-to-date on microorganisms in foods. Topics such as DNA sequencing and E. coli are particularly well covered. With lists of further reading to help users explore topics in depth, this resource will enrich scientists at every level in academia and industry, providing fundamental information as well as explaining state-of-the-art scientific discoveries. This book is designed to allow disparate approaches (from farmers to processors to food handlers and consumers) and interests to access accurate and objective information about the microbiology of foods Microbiology impacts the safe presentation of food. From harvest and storage to determination of shelf-life, to presentation and consumption. This work highlights the risks of microbial contamination and is an invaluable go-to guide for anyone working in Food Health and Safety Has a two-fold industry appeal (1) those developing new functional food products and (2) to all corporations concerned about the potential hazards of microbes in their food products
