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JoVE Science Education

General Laboratory Techniques

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Basic Methods in Cellular and Molecular Biology

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Essentials of Biology 1

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JoVE Biology
JoVE Biology welcomes all general biology research methodologies. Content in this section canvases all fields of cell, molecular, and organismal biology, ranging from new applications of standard techniques to novel approaches aimed at understanding the functions of life and living organisms. This diverse section includes, but is not limited to, techniques in physical biology, cellular biochemistry, genetics, physiology, systems biology and a combination of eukaryotic and prokaryotic model systems.
 JoVE Biology

A Protocol for Lentiviral Transduction and Downstream Analysis of Intestinal Organoids

1Tytgat Institute for Liver and Intestinal Research, Department of Gasteroenterology and Hepatology, Academical Medical Center


JoVE 52531

In this video protocol we give a step by step explanation of lentiviral transduction in organoids of primary intestinal epithelium and of processing and downstream analysis of these cultures by quantitative RT-PCR, RNA-microarray and immunohistochemistry.

 JoVE Biology

Determining Membrane Protein Topology Using Fluorescence Protease Protection (FPP)

1Department of Physiology and Biophysics, Chicago Medical School


JoVE 52509

Here, we present a protocol to determine the orientation and topology of integral membrane proteins in living cells. This simple protocol relies on selective protease sensitivity of chimeras between the protein of interest and GFP.

 JoVE Biology

A Proteoliposome-Based Efflux Assay to Determine Single-molecule Properties of Cl- Channels and Transporters

1Department of Anesthesiology, Weill Cornell Medical College, 2Department of Physiology and Biophysics, Weill Cornell Medical College, 3Department of Biochemistry, Weill Cornell Medical College


JoVE 52369

Proteoliposomes are used to study purified channels and transporters reconstituted in a well-defined biochemical environment. An experimental procedure to measure efflux mediated by these proteins is illustrated. The steps to prepare proteoliposomes, perform the recordings, and analyze data to quantitatively determine the functional properties of the reconstituted protein are described.

 JoVE Biology

Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs

1Diabetes and Islet Biology Group, NHMRC Clinical Trials Centre, Faculty of Medicine, The University of Sydney, 2Biomarkers Laboratory, NHMRC Clinical Trials Centre, Faculty of Medicine, The University of Sydney


JoVE 52586

Circulating microRNAs have recently emerged as promising and novel biomarkers for various cancers and other diseases. The goal of this article is to discuss three different probe-based real-time PCR platforms and methods that are available to quantify and determine the abundance of circulating microRNAs.

 JoVE Biology

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications

1Department of Biology, San Diego State University


JoVE 52452

Since the discovery of the green fluorescent protein gene, fluorescent proteins have impacted molecular cell biology. This protocol describes how expression of distinct fluorescent proteins through genetic engineering is used for barcoding individual cells. The procedure enables tracking distinct populations in a cell mixture, which is ideal for multiplexed applications.

 JoVE Biology

RNA Isolation from Cell Specific Subpopulations Using Laser-capture Microdissection Combined with Rapid Immunolabeling

1Department of Psychiatry and Neurosciences, Faculty of Medicine, Université Laval, 2Centre de recherche de l'Institut universitaire en santé mentale de Québec


JoVE 52510

Gene expression analysis of a subset of cells in a specific tissue represents a major challenge. This article describes how to isolate high-quality total RNA from a specific cell population by combining a rapid immunolabeling method with laser capture microdissection.

 JoVE Biology

Synchronization of Caulobacter Crescentus for Investigation of the Bacterial Cell Cycle

1Department of Developmental Biology, Stanford University School of Medicine


JoVE 52633

Synchronization of bacterial cells is essential for studies of the bacterial cell cycle and development. Caulobacter crescentus is synchronizable through density centrifugation allowing a rapid and powerful tool for studies of the bacterial cell cycle. Here we provide a detailed protocol for the synchronization of Caulobacter cells.

 JoVE Biology

Preparation, Imaging, and Quantification of Bacterial Surface Motility Assays

1Department of Civil and Environmental Engineering and Earth Sciences, University of Notre Dame, 2Eck Institute for Global Health, University of Notre Dame, 3Department of Applied and Computational Mathematics and Statistics, University of Notre Dame, 4INRS-Institut Armand-Frappier, 5Department of Biology, Indiana University, 6Department of Biological Sciences, University of Notre Dame


JoVE 52338

Swarming motility is influenced by physical and environmental factors. We describe a two-phase protocol and guidelines to circumvent the challenges commonly associated with swarm assay preparation and data collection. A macroscopic imaging technique is employed to obtain detailed information on swarm behavior that is not provided by current analysis techniques.

 JoVE Biology

Rapid Identification of Chemical Genetic Interactions in Saccharomyces cerevisiae

1Department of Biochemistry and Microbiology, University of Victoria


JoVE 52345

Here we present a cost-effective method for defining chemical-genetic interactions in budding yeast. The approach is built on fundamental techniques in yeast molecular biology and is well suited for the mechanistic interrogation of small to medium collections of chemicals and other media environments.

 JoVE Biology

Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control

1Department of Bioengineering, Rice University


JoVE 52620

Provided is a protocol for developing a real-time recombinase polymerase amplification assay to quantify initial concentration of DNA samples using either a thermal cycler or a microscope and stage heater. Also described is the development of an internal positive control. Scripts are provided for processing raw real-time fluorescence data.

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