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National Research Council Canada

7 ARTICLES PUBLISHED IN JoVE

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Biology

Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons
Sylvie Sanschagrin 1, Etienne Yergeau 1
1Energy, Mining and Environment, National Research Council Canada

Characterizing microbial community has been a longstanding goal in environmental microbiology. Next-generation sequencing methods now allow for the characterization of microbial communities at an unprecedented depth with minimal cost and labor. We detail here our approach to sequence bacterial 16S ribosomal RNA genes using a benchtop sequencer.

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Immunology and Infection

Isolation and Characterization of Extracellular Vesicles from Adult Schistosoma japonicum
Juntao Liu *1, Lihui Zhu *1, Lihui Wang 1,2, Yongjun Chen 1, Bikash Ranjan Giri 1, Jianjun Li 2, Guofeng Cheng 1
1Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology, Ministry of Agriculture, 2Tianjin Agricultural University

Here, we present a protocol to describe extracellular vesicles (EVs) isolation in in vitro cultured medium from adult Schistosoma japonicum.

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JoVE Core

Intermediate Strain Rate Material Characterization with Digital Image Correlation
Meysam Rahmat 1, David Backman 1, Richard Desnoyers 1
1Aerospace Research Centre, National Research Council Canada

Here we present a methodology for the dynamic characterization of tensile specimens at intermediate strain rates using a high-speed servo-hydraulic load frame. Procedures for strain gauge instrumentation and analysis, as well as for digital image correlation strain measurements on the specimens, are also defined.

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Neuroscience

Stimulation Location Determination using a 3D Digitizer with High-Definition Transcranial Direct Current Stimulation
Wanting Chen 1, Rui Chen 1, Qinghua He 1,2,3,4,5
1Faculty of Psychology, Southwest University, 2Key Laboratory of Cognition and Personality, Ministry of Education, Southwest University, 3Southwest University Branch, Collaborative Innovation Center of Assessment toward Basic Education Quality, Beijing Normal University, 4Key Laboratory of Mental Health, Institute of Psychology, Chinese Academy of Sciences, 5Chongqing Collaborative Innovation Center for Brain Science

Presented here is a protocol to achieve higher accuracy in determination of stimulation location combining a 3D digitizer with high-definition transcranial direct current stimulation.

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Bioengineering

Fabrication of a Crystalline Nanocellulose Embedded Agarose Biomaterial Ink for Bone Marrow-Derived Mast Cell Culture
Leshern Karamchand 1, Ashley Wagner 1, Syed Benazir Alam 1, Marianna Kulka 1,2
1Nanotechnology Research Center, National Research Council Canada, 2Department of Medical Microbiology and Immunology, University of Alberta

This protocol highlights a method to rapidly assess the biocompatibility of a crystalline nanocellulose (CNC)/agarose composite hydrogel biomaterial ink with mouse bone marrow-derived mast cells in terms of cell viability and phenotypic expression of the cell surface receptors, Kit (CD117) and high-affinity IgE receptor (FcεRI).

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Immunology and Infection

Generation of Null Mutants to Elucidate the Role of Bacterial Glycosyltransferases in Bacterial Motility
Juan M. Tomás 1, Kelly M. Fulton 2,3, Susan M. Twine 2,4, Susana Merino 1
1Departamento de Genética, Microbiología y Estadística, Universidad de Barcelona, 2Human Health Therapeutics Research Centre, National Research Council Canada, 3Department of Chemistry, Carleton University, 4Department of Biology, Carleton University

Here, we describe the construction of null mutants of Aeromonas in specific glycosyltransferases or regions containing glycosyltransferases, the motility assays, and flagella purification performed to establish the involvement and function of their encoded enzymes in the biosynthesis of a glycan, as well as the role of this glycan in bacterial pathogenesis.

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Biochemistry

Nonspecific Proteolysis-Based Glycomics Strategy for Bacterial Glycoproteins
Rui Chen 1, Jacek Stupak 1, Jianjun Li 1
1Human Health Therapeutics Research Center, National Research Council Canada

This study presents a method for glycomics analysis of glycoproteins by a combination of pronase E digestion, permethylation, and mass spectrometry analysis. This method is capable of analyzing all types of N-linked glycans, including bacterial N-glycans.

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