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Detection and Genogrouping of Noroviruses from Children's Stools By Taqman One-step RT-PCR

1Laboratorio de Investigación y Desarrollo (LID), Universidad Peruana Cayetano Heredia, 2Bloomberg School of Public Health, Johns Hopkins University, 3Laboratorio de Diagnostico Molecular, Facultad de Medicina, University of Concepcion,Chile, 4University of California San Diego School of Medicine

JoVE 3232


 Medicine

Developing a Salivary Antibody Multiplex Immunoassay to Measure Human Exposure to Environmental Pathogens

1National Exposure Research Laboratory, U.S. Environmental Protection Agency, 2National Risk Management Research Laboratory, U.S. Environmental Protection Agency, 3Oak Ridge Institute for Science and Education, 4Department of Biological Sciences, McMicken College of Arts and Sciences, University of Cincinnati

JoVE 54415


 Immunology and Infection

RNA Analysis of Environmental Samples Using RT-PCR

JoVE 10104

Source: Laboratories of Dr. Ian Pepper and Dr. Charles Gerba - Arizona University
Demonstrating Author: Bradley Schmitz

Reverse transcription-polymerase chain reaction (RT-PCR) involves the same process as conventional PCR — cycling temperature to amplify nucleic acids. However, while conventional PCR only amplifies deoxyribonucleic acids (DNA), RT-PCR enables the amplification of ribonucleic acids (RNA) through the formation of complementary DNA (cDNA). This enables RNA-based organisms found within the environment to be analyzed utilizing methods and technologies that are designed for DNA. Many viruses found in the environment use RNA as their genetic material. Several RNA-based viral pathogens, such as Norovirus, and indicator organisms, such as pepper mild mottle virus (PMMoV), do not have culture-based detection methods for quantification. In order to detect for the presence of these RNA viruses in environmental samples from soil, water, agriculture, etc., molecular assays rely on RT-PCR to convert RNA into DNA. Without RT-PCR, microbiologists would not be able to assay and research numerous RNA-based viruses that pose risks to human and environmental health. RT-PCR can also be employed as a tool to measure microbial activity in the env


 Environmental Microbiology

Breast Milk Enhances Growth of Enteroids: An Ex Vivo Model of Cell Proliferation

1Division of Newborn Medicine, Department of Pediatrics, Washington University School of Medicine, 2Washington University, 3Division of Newborn Medicine, Department of Pediatrics, University of Pittsburgh School of Medicine

Video Coming Soon

JoVE 56921


 JoVE In-Press

Quantifying Environmental Microorganisms and Viruses Using qPCR

JoVE 10186

Source: Laboratories of Dr. Ian Pepper and Dr. Charles Gerba - Arizona University
Demonstrating Author: Bradley Schmitz

Quantitative polymerase chain reaction (qPCR), also known as real-time PCR, is a widely-used molecular technique for enumerating microorganisms in the environment. Prior to this approach, quantifying microorganisms was limited largely to classical culture-based techniques. However, the culturing of microbes from environmental samples can be particularly challenging, and it is generally held that as few as 1 to 10% of the microorganisms present within environmental samples are detectable using these techniques. The advent of qPCR in environmental microbiology research has therefore advanced the field greatly by allowing for more accurate determination of concentrations of microorganisms such as disease-causing pathogens in environmental samples. However, an important limitation of qPCR as an applied microbiological technique is that living, viable populations cannot be differentiated from inactive or non-living populations. This video demonstrates the use of qPCR to detect pepper mild mottle virus from an environmental water sample.


 Environmental Microbiology

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Streamlined Single Cell TCR Isolation and Generation of Retroviral Vectors for In Vitro and In Vivo Expression of Human TCRs

1Department of Pediatrics, Section of Diabetes and Endocrinology, McNair Medical Institute, Baylor College of Medicine, Texas Children's Hospital, 2Benaroya Research Institute at Virginia Mason, 3Center for Human Immunobiology, Baylor College of Medicine, Texas Children's Hospital, 4Department of Pediatrics, Section of Diabetes and Endocrinology, Baylor College of Medicine, Texas Children's Hospital

JoVE 55379


 Immunology and Infection

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High Efficiency Differentiation of Human Pluripotent Stem Cells to Cardiomyocytes and Characterization by Flow Cytometry

1Department of Biochemistry, Medical College of Wisconsin, 2Stanford Cardiovascular Institute, Stanford University School of Medicine, 3Department of Anesthesiology, Medical College of Wisconsin, 4Stem Cell and Regenerative Medicine Consortium, LKS Faculty of Medicine, Hong Kong University, 5Division of Cardiology, Johns Hopkins University School of Medicine, 6Cardiovascular Research Center, Biotechnology and Bioengineering Center, Medical College of Wisconsin

JoVE 52010


 Biology

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Chemically-blocked Antibody Microarray for Multiplexed High-throughput Profiling of Specific Protein Glycosylation in Complex Samples

1Institute for Hepatitis and Virus Research, 2Department of Microbiology and Immunology, Thomas Jefferson University, 3Drexel University College of Medicine, 4Van Andel Research Institute, 5Institute for Hepatitis and Virus Research, Serome Biosciences Inc.

JoVE 3791


 Biology

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Optimized Protocol for the Extraction of Proteins from the Human Mitral Valve

1Centro Cardiologico Monzino IRCCS, 2Cardiovascular Tissue Bank of Milan, Centro Cardiologico Monzino IRCCS, 3Department of Clinical Sciences and Community Health, Cardiovascular Section, University of Milan, 4Department of Cardiovascular Disease, Development and Innovation Cardiac Surgery Unit, Centro Cardiologico Monzino IRCCS

JoVE 55762


 Biochemistry

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Rapid Neuronal Differentiation of Induced Pluripotent Stem Cells for Measuring Network Activity on Micro-electrode Arrays

1Department of Cognitive Neurosciences, Radboudumc, 2Donders Institute for Brain, Cognition and Behaviour, Radboud University, 3Department of Human Genetics, Radboudumc, 4Department of Molecular Developmental Biology, Radboud University

JoVE 54900


 Developmental Biology

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Contractility Measurements of Human Uterine Smooth Muscle to Aid Drug Development

1Harris-Wellbeing Preterm Birth Research Centre, Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, 2School of Biomedical Sciences, The University of Queensland, 3Institute for Molecular Bioscience, The University of Queensland, 4Center for Physiology and Pharmacology, Medical University of Vienna

Video Coming Soon

JoVE 56639


 JoVE In-Press

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Reprogramming Primary Amniotic Fluid and Membrane Cells to Pluripotency in Xeno-Free Conditions

1Mitchell Cancer Institute, University of South Alabama, 2College of Medicine, University of South Alabama, 3Institute for Regenerative Medicine, University of Zurich, 4Department of Dermatology, University Hospital Zurich, 5Center for Applied Biotechnology and Molecular Medicine (CABMM), University of Zurich - Irchel Campus

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JoVE 56003


 JoVE In-Press

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Human Pluripotent Stem Cell Based Developmental Toxicity Assays for Chemical Safety Screening and Systems Biology Data Generation

1Center of Physiology and Pathophysiology, Institute of Neurophysiology, University of Cologne, 2Department of Biology, University of Konstanz, 3Department of Statistics, Technical University of Dortmund, 4Leibniz Research Centre for Working Environment and Human Factors, Technical University of Dortmund

JoVE 52333


 Developmental Biology

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Chondrogenic Pellet Formation from Cord Blood-derived Induced Pluripotent Stem Cells

1CiSTEM Laboratory, Convergent Research Consortium for Immunologic Disease, Division of Rheumatology, Seoul St. Mary's Hospital, College of Medicine, The Catholic University of Korea, 2Division of Rheumatology, Department of Internal Medicine, Seoul St. Mary's Hospital, Institute of Medical Science, College of Medicine, The Catholic University of Korea

JoVE 55988


 Developmental Biology

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