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General Laboratory Techniques

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Basic Methods in Cellular and Molecular Biology

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 JoVE General

Preparation of Quality Inositol Pyrophosphates

1Medical Research Council (MRC), Cell Biology Unit and Laboratory for Molecular Cell Biology, University College London


JoVE 3027

Inositol pyrophosphates play an important role in human pathologies such cancer, diabetes and obesity; however, the exact mechanism of action is a matter of dispute. The lack of commercially available inositol pyrophosphates renders detailed studies problematic. Here we describe a simple protocol to produce and isolate milligrams of inositol pyrophosphates.

 JoVE Immunology and Infection

Expansion of Human Peripheral Blood γδ T Cells using Zoledronate

1Department of Immunotherapeutics (Medinet), University of Tokyo Hospital, 2MEDINET Co., Ltd


JoVE 3182

A method to expand γδ T cells from peripheral blood mononuclear cells (PBMC) is described. PBMC-derived γδ T cells are stimulated and expanded using zoledronate and interleukin-2 (IL-2). Large scale expansion of γδ T cells can be applied to autologous cellular immunotherapy of cancer.

 JoVE Chemistry

Nucleoside Triphosphates - From Synthesis to Biochemical Characterization

1Department of Chemistry and Biochemistry, University of Bern


JoVE 51385

The protocol described herein aims to explain and abridge the numerous obstacles in the way of the intricate route leading to modified nucleoside triphosphates. Consequently, this protocol facilitates both the synthesis of these activated building-blocks and their availability for practical applications.

 JoVE General

Pyrosequencing for Microbial Identification and Characterization

1Center for Biotechnology Education, Krieger School of Arts and Sciences, Johns Hopkins University, 2Qiagen Sciences, Inc.


JoVE 50405

Pyrosequencing is a versatile technique that facilitates microbial genome sequencing that can be used to identify bacterial species, discriminate bacterial strains, and detect genetic mutations that confer resistance to anti-microbial agents. In this video, the procedure for microbial amplicon generation, amplicon pyrosequencing, and DNA sequence analysis will be demonstrated.

 JoVE General

Direct Detection of the Acetate-forming Activity of the Enzyme Acetate Kinase

1Department of Genetics and Biochemistry, Clemson University


JoVE 3474

A method for the determination of acetate kinase activity is described. This assay utilizes a direct reaction for determining enzyme activity and kinetics of acetate kinase in the acetate-forming direction with different phosphoryl acceptors. Furthermore, this method can be utilized for assaying other acetyl phosphate or acetyl-CoA utilizing enzymes.

 JoVE Immunology and Infection

Use of Galleria mellonella as a Model Organism to Study Legionella pneumophila Infection

1Center for Molecular Bacteriology and Infection, Imperial College London


JoVE 50964

The larva of the wax moth Galleria mellonella was recently established as an in vivo model to study Legionella pneumophila infection. Here, we demonstrate fundamental techniques to characterize the pathogenesis of Legionella in the larvae, including inoculation, measurement of bacterial virulence and replication as well as extraction and analysis of infected hemocytes.

 JoVE Immunology and Infection

In vitro tRNA Methylation Assay with the Entamoeba histolytica DNA and tRNA Methyltransferase Dnmt2 (Ehmeth) Enzyme

1Faculty of Medicine, Rappaport Institute, Technion - Israel Institute of Technology, 2The Pharmacy and Biochemistry Institute, Johannes Gutenberg University


JoVE 2390

This protocol describes the preparation of a synthetic tRNA substrate for the Entamoeba histolytica DNA/tRNA methyltransferase 2 (Dnmt2) homolog Ehmeth and the measure of its methyltransferase activity. This experimental approach can be used for investigating the activity of other Dnmt2 proteins.

 JoVE Bioengineering

Hydrophobic Salt-modified Nafion for Enzyme Immobilization and Stabilization

1Departments of Chemistry and Materials Science and Engineering, University of Utah


JoVE 3949

This article will describe the procedure for synthesizing a hydrophobically modified Nafion enzyme immobilization membrane and how to immobilize proteins and/or enzymes within the membrane and test their specific activity.

 JoVE General

Method for the Isolation of Francisella tularensis Outer Membranes

1Department of Microbiology, University of Texas Southwestern Medical Center


JoVE 2044

A protocol for separating inner and outer membranes from Francisella tularensis by spheroplasting, osmotic lysis, and sucrose density gradient ultracentrifugation.

 JoVE General

Analyzing Gene Expression from Marine Microbial Communities using Environmental Transcriptomics

1Department of Marine Sciences, University of Georgia (UGA)


JoVE 1086

We present a method for generating cDNA from environmental mRNA. In general, total RNA is first collected from the environment, rRNA is selectively removed, mRNA is selectively amplified, and cDNA synthesized from the enriched mRNA pool is sequenced. Recovered sequences can be annotated using standard bioinformatics techniques to identify the expressed genes.

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