Download or read book Reporter Gene Assays written by Robert Damoiseaux and published by Humana Press. This book was released on 2018-04-19 with total page 234 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume explores the applications of reporter gene technology and the methodologies needed for their effective implementation. The chapters in this book cover practical topics such as how to integrate reporter constructs into cellular models, viral delivery, splicing applications, in vivo imaging, and a guide to the use of multi-cistronic constructs. Additionally, chapters also include detailed mechanistic uses of reporter genes in cellular pathways, and a look at project and data management of screening applications. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions 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 thorough, Reporter Gene Assays: Methods and Protocols is a valuable resource for anyone who is interested in learning more about reporter genes.
Download or read book Encyclopedia of Cancer written by Manfred Schwab and published by Springer Science & Business Media. This book was released on 2008-09-23 with total page 3307 pages. Available in PDF, EPUB and Kindle. Book excerpt: This comprehensive encyclopedic reference provides rapid access to focused information on topics of cancer research for clinicians, research scientists and advanced students. Given the overwhelming success of the first edition, which appeared in 2001, and fast development in the different fields of cancer research, it has been decided to publish a second fully revised and expanded edition. With an A-Z format of over 7,000 entries, more than 1,000 contributing authors provide a complete reference to cancer. The merging of different basic and clinical scientific disciplines towards the common goal of fighting cancer makes such a comprehensive reference source all the more timely.
Download or read book Molecular Diagnostics Promises and Possibilities written by Mousumi Debnath and published by Springer Science & Business Media. This book was released on 2010-01-29 with total page 527 pages. Available in PDF, EPUB and Kindle. Book excerpt: A rapid development in diverse areas of molecular biology and genetic engineering resulted in emergence of variety of tools. These tools are not only applicable to basic researches being carried out world over, but also exploited for precise detection of abnormal conditions in plants, animals and human body. Although a basic researcher is well versed with few techniques used by him/her in the laboratory, they may not be well acquainted with methodologies, which can be used to work out some of their own research problems. The picture is more blurred when the molecular diagnostic tools are to be used by physicians, scientists and technicians working in diagnostic laboratories in hospitals, industry and academic institutions. Since many of them are not trained in basics of these methods, they come across several gray areas in understanding of these tools. The accurate application of molecular diagnostic tools demands in depth understanding of the methodology for precise detection of the abnormal condition of living body. To meet the requirements of a good book on molecular diagnostics of students, physicians, scientists working in agricultural, veterinary, medical and pharmaceutical sciences, it needs to expose the reader lucidly to: Give basic science behind commonly used tools in diagnostics Expose the readers to detailed applications of these tools and Make them aware the availability of such diagnostic tools The book will attract additional audience of pathologists, medical microbiologists, pharmaceutical sciences, agricultural scientists and veterinary doctors if the following topics are incorporated at appropriate places in Unit II or separately as a part of Unit-III in the book. Molecular diagnosis of diseases in agricultural crops Molecular diagnosis of veterinary diseases. Molecular epidemiology, which helps to differentiate various epidemic strains and sources of disease outbreaks. Even in different units of the same hospital, the infections could be by different strains of the same species and the information becomes valuable for infection control strategies. Drug resistance is a growing problem for bacterial, fungal and parasitic microbes and the molecular biology tools can help to detect the drug resistance genes without the cultivation and in vitro sensitivity testing. Molecular diagnostics offers faster help in the selection of the proper antibiotic for the treatment of tuberculosis, which is a major problem of the in the developing world. The conventional culture and drug sensitivity testing of tuberculosis bacilli is laborious and time consuming, whereas molecular diagnosis offers rapid drug resistant gene detection even from direct clinical samples. The same approach for HIV, malaria and many more diseases needs to be considered. Molecular diagnostics in the detection of diseases during foetal life is an upcoming area in the foetal medicine in case of genetic abnormalities and infectious like TORCH complex etc. The book will be equally useful to students, scientists and professionals working in the field of molecular diagnostics.
Download or read book GUS Protocols written by Sean R. Gallagher and published by Academic Press. This book was released on 2012-12-02 with total page 250 pages. Available in PDF, EPUB and Kindle. Book excerpt: The gb-glucuronidase (GUS) gene is extremely useful as a reporter of the expression of introduced genes and can be used in organisms where other reporter genes are useless. Thus, the GUS gene is the reporter gene of choice for transgenic plant research. Not only can this assay be used to detect whether a gene is being expressed, but it can be used to determine the location of the gene product within cells. Low cost is another advantage of this assay. GUS Protocols provides instructions and essential background information that will enable researchers to effectively use the GUS gene as a reporter of the expression of introduced genes. First book on the GUS reporter system Up-to-date reference lists following each of the fourteen chapters Comb-bound for convenient bench-top use Written by leading authorities including R.A. Jefferson, inventor of the GUS assay Illustrated with color GUS detection by fluorometric, spectrophotometric, and histochemical methods Fast, automated assays
Download or read book Test No 488 Transgenic Rodent Somatic and Germ Cell Gene Mutation Assays written by OECD and published by OECD Publishing. This book was released on 2011-07-28 with total page 16 pages. Available in PDF, EPUB and Kindle. Book excerpt: This Test Guideline describes an in vivo assay that detects chemicals that may induce gene mutations. In this assay, transgenic rats or mice that contain multiple copies of chromosomally integrated plasmid or phage shuttle vectors are used. The ...
Download or read book In Vitro 3D Fluorescent Cell based Assay Reporting Gene Regulation for High throughput Drug Screening written by You Li and published by . This book was released on 2022 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Traditional cell-based assays have a high failure rate of identifying effective compounds in drug discovery. To address this problem, recent drug research has focused on engineering cancer cells that can indicate specific drug molecular mechanisms of action and developing cancer models that can more accurately represent tumor physiology. Survivin and telomerase reverse transcriptase (TERT), two important cancer biomarkers overexpressed in 85% of tumor tissues, play crucial roles in anti-apoptosis and unlimited cell proliferation in carcinogenesis, as well as drug resistance in cancer therapy. In this research, we have developed survivin and TERT reporter gene assays for high-throughput screening (HTS) of drugs downregulating these genes as anticancer therapeutics. Cancer cells were engineered to express fluorescent proteins controlled by TERT and survivin promoters, respectively, and were cultured in 3-dimensional (3D) scaffolds in microbioreactors to investigate drug effects by monitoring changes in culture fluorescence in real time. The novel 3-D assays offered molecular and physiological predictions of gene-specific responses to drugs, thus facilitating cancer drug discovery and development. Using survivin as the target, a dual-color fluorescent reporter gene assay performed in 3-D culture was developed. To demonstrate the drug's specific effect on the survivin promoter, MCF-7 breast cancer cells were engineered to express enhanced green fluorescent protein (EGFP) driven by a survivin promoter. The cell also expressed red fluorescent protein (RFP) under the control of a cytomegalovirus promoter as an internal control and to indicate cell number. The specific survivin expression was assessed by the green fluorescence normalized to the red fluorescence, which was used as an indicator to assess drug’s regulation on survivin. The internal control compensated for the variations in cell number, cell culture environment, fluorescent protein property, detection setting, and even for the reading interference from tested compounds, resulting in a more accurate evaluation of drug effect. The assay correctly detected the downregulation and upregulation of several anticancer compounds and demonstrated its reliability for high-throughput screening of survivin-targeted drugs. In this work, a 3-D cancer cell-based hTERT promoter assay was also developed for anticancer drug screening. Different lengths of the hTERT promoter were cloned with EGFP as reporting cassettes and were introduced in multiple cancer cell lines. The selected MCF7 reporter cell was able to correctly respond to the known hTERT inhibitors, pristimerin, epigallocatechin gallate and n-butylidenephthalide. The reporter cells were cultured on 3-D scaffolds to resemble in vivo tumor physiology and present in vivo like responses. The 3-D hTERT promoter assay was verified by pristimerin, and subsequently, other five anticancer compounds, including doxorubicin, cisplatin, paclitaxel, blasticidin, and tamoxifen, were tested for their effects on hTERT. The developed assay achieved an overall accuracy rate of over 80%, which demonstrated the feasibility of using 3D culture and EGFP as a reporter to screen hTERT inhibitors. In addition to the development of cancer cell-based assays, hTERT as a pluripotency marker to predict embryotoxic compounds during tissue development was also evaluated. The hTERT-EGFP reporting cassettes were introduced into mouse embryonic stem cells D3. The reportability was assessed by the corelated expression changes of EGFP and endogenous TERT during neural differentiation and after receiving toxicant stimuli. Our results showed that EGFP was able to indicate drug effect on human TERT expression level during ESC differentiation, which, however, was not always consistent with internal mouse TERT (mTERT) due to species difference. The results confirmed that drug effects on TERT were species-specific, especially at the beginning of differentiation. In conclusion, hTERT in mESCs reacted to embryotoxic compounds and the engineered mESC-hTERT-EGFP cells could be used for human teratogen prediction.
Download or read book A Study of Reporter Gene Assays for the Analysis of Gene Expression written by and published by . This book was released on 1992 with total page 814 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Guide to Research Techniques in Neuroscience written by Matt Carter and published by Academic Press. This book was released on 2022-03-26 with total page 416 pages. Available in PDF, EPUB and Kindle. Book excerpt: Modern neuroscience research is inherently multidisciplinary, with a wide variety of cutting edge new techniques to explore multiple levels of investigation. This Third Edition of Guide to Research Techniques in Neuroscience provides a comprehensive overview of classical and cutting edge methods including their utility, limitations, and how data are presented in the literature. This book can be used as an introduction to neuroscience techniques for anyone new to the field or as a reference for any neuroscientist while reading papers or attending talks. Nearly 200 updated full-color illustrations to clearly convey the theory and practice of neuroscience methods Expands on techniques from previous editions and covers many new techniques including in vivo calcium imaging, fiber photometry, RNA-Seq, brain spheroids, CRISPR-Cas9 genome editing, and more Clear, straightforward explanations of each technique for anyone new to the field A broad scope of methods, from noninvasive brain imaging in human subjects, to electrophysiology in animal models, to recombinant DNA technology in test tubes, to transfection of neurons in cell culture Detailed recommendations on where to find protocols and other resources for specific techniques "Walk-through" boxes that guide readers through experiments step-by-step
Download or read book E coli Plasmid Vectors written by Nicola Casali and published by Humana Press. This book was released on 2010-11-10 with total page 316 pages. Available in PDF, EPUB and Kindle. Book excerpt: A comprehensive collection of readily reproducible techniques for the manipulation of recombinant plasmids using the bacterial host E. coli. The authors describe proven methods for cloning DNA into plasmid vectors, transforming plasmids into E. coli, and analyzing recombinant clones. They also include protocols for the construction and screening of libraries, as well as specific techniques for specialized cloning vehicles, such as cosmids, bacterial artificial chromosomes, l vectors, and phagemids. Common downstream applications such as mutagenesis of plasmids, recombinant protein expression, and the use of reporter genes, are also described.
Download or read book ERE and XRE based Reporter Gene Assays to Detect Biological Effects of Environmental Chemicals written by Do-Minh-Si and published by . This book was released on 2007 with total page 358 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Molecular Biology and Genomics written by Cornel Mulhardt and published by Elsevier. This book was released on 2010-07-19 with total page 273 pages. Available in PDF, EPUB and Kindle. Book excerpt: Never before has it been so critical for lab workers to possess the proper tools and methodologies necessary to determine the structure, function, and expression of the corresponding proteins encoded in the genome. Mulhardt's Molecular Biology and Genomics helps aid in this daunting task by providing the reader with tips and tricks for more successful lab experiments. This strategic lab guide explores the current methodological variety of molecular biology and genomics in a simple manner, addressing the assets and drawbacks as well as critical points. It also provides short and precise summaries of routine procedures as well as listings of the advantages and disadvantages of alternative methods. Shows how to avoid experimental dead ends and develops an instinct for the right experiment at the right time Includes a handy Career Guide for researchers in the field Contains more than 100 extensive figures and tables
Download or read book Small Non Coding RNAs written by Mathieu Rederstorff 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 state-of-the-art methods tackling all aspects of small non-coding RNAs biology. Small Non-Coding RNAs: Methods and Protocols guides readers through customized dedicated protocols and technologies that will be of valuable help to all those willing to contribute deciphering the numerous functions of small non-coding RNAs. Written in the highly successful Methods of Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols and key tips on troubles troubleshooting and avoiding known pitfalls. Instructive and practical, Small Non-Coding RNAs: Methods and Protocols reaches out to biochemists, cellular and molecular biologists already working in the field of RNA biology and to those just starting to study small non-coding RNAs.
Download or read book Gene Regulatory Sequences and Human Disease written by Nadav Ahituv and published by Springer Science & Business Media. This book was released on 2012-05-30 with total page 289 pages. Available in PDF, EPUB and Kindle. Book excerpt: In Gene Regulatory Sequences and Human Disease, the Editor will introduce the different technological advances that led to this breakthrough. In addition, several examples will be provided of nucleotide variants in noncoding sequences that have been shown to be associated with various human diseases.
Download or read book Disease Gene Identification written by Johanna K. DiStefano and published by Humana. This book was released on 2019-06-06 with total page 400 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume presents detailed laboratory procedures in an easy to follow format that can be carried out with success by investigators lacking previous exposure to a specific research method. Chapter guide readers through the application of molecular approaches to disease gene identification and overviews, and case studies are also presented. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions 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. Authoritative and practical, Disease Gene Identification: Methods and Protocols, Second Edition aims to help with the identification and characterization of many more disease-related genes and provide novel, and effective strategies for disease treatment and prevention.
Download or read book HIV Protocols written by Vinayaka R. Prasad and published by Humana Press. This book was released on 2008-12-08 with total page 457 pages. Available in PDF, EPUB and Kindle. Book excerpt: Despite major advances in HIV treatment, many areas require more study, in order to create efficacious, potent antiretrovirals that can suppress viral load completely and durably without toxic side effects, to define unknown drug targets and fine-tune known targets, and to better understand the interplay between viral and host factors. In "HIV Protocols, Second Edition", expert researchers provide clear, state-of-the-art methods for the study of HIV. Directed toward three specific goals, this text aims to document up-to-date protocols for select aspects of HIV biology, to bring together both virological and immunological approaches in a single, convenient volume, and to present a comprehensive account of a range of techniques not available in any existing HIV protocol book. As a volume in the highly successful Methods in Molecular BiologyTM series, the chapters include brief introductions to the subject, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and Notes sections containing priceless tips on troubleshooting and avoiding known pitfalls. Comprehensive and cutting-edge, "HIV Protocols, Second Edition" is an ideal guide to the wide array of techniques used in fundamental or applied research into the biology and pathogenesis of HIV-1.
Download or read book Assay Development written by Ge Wu and published by John Wiley & Sons. This book was released on 2010-04-26 with total page 450 pages. Available in PDF, EPUB and Kindle. Book excerpt: Essential principles and practice of assay development The first comprehensive, integrated treatment of the subject, Assay Development: Fundamentals and Practices covers the essentials and techniques involved in carrying out an assay project in either a biotechnology/drug discovery setting or a platform setting. Rather than attempting comprehensive coverage of all assay development technologies, the book introduces the most widely used assay development technologies and illustrates the art of assay development through a few commonly encountered biological targets in assay development (e.g., proteases, kinases, ion channels, and G protein-coupled receptors). Just enough biological background for these biological targets is provided so that the reader can follow the logics of assay development. Chapters discuss: The basics of assay development, including foundational concepts and applications Commonly used instrumental methods for both biochemical assays and cell-based assays Assay strategies for protein binding and enzymatic activity Cell-based assays High-throughput screening An in-depth study of the now popular Caliper's off-chip kinase assay provides an instructive, real-world example of the assay development process.
Download or read book Chemical Genomics written by Haian Fu and published by Cambridge University Press. This book was released on 2012-02-13 with total page 359 pages. Available in PDF, EPUB and Kindle. Book excerpt: Advances in chemistry, biology and genomics coupled with laboratory automation and computational technologies have led to the rapid emergence of the multidisciplinary field of chemical genomics. This edited text, with contributions from experts in the field, discusses the new techniques and applications that help further the study of chemical genomics. The beginning chapters provide an overview of the basic principles of chemical biology and chemical genomics. This is followed by a technical section that describes the sources of small-molecule chemicals; the basics of high-throughput screening technologies; and various bioassays for biochemical-, cellular- and organism-based screens. The final chapters connect the chemical genomics field with personalized medicine and the druggable genome for future discovery of new therapeutics. This book will be valuable to researchers, professionals and graduate students in many fields, including biology, biomedicine and chemistry.