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Protocols

protocols
Protocol

Varicella Zoster Virus Replication Assays

Experiment Tags: Varicella Zoster Virus (VZV), Replication Assays

Experiment Summary: This protocol describes a basic VZV replication assay using a recombinant VZV-GFP reporter virus. This reporter virus-infected cell line can be used in combination with siRNA gene depletion or cDNA overexpression transfection protocols to determine the effect of individual cellular genes on virus replication.

protocols
Protocol

Protocol for Rate-Controlled Cryopreservation and Thawing of Mammalian Cells

Experiment Tags: Biological Techniques, Cryopreservation, Cooling, Freezing

Experiment Summary: In this protocol, we describe the general procedure for cryopreservation of cultured cells.

protocols
Protocol

Protocol for Intrathoracic Inoculation of Zika Virus in Aedes aegypti

Experiment Tags: Aedes aegypti, Mosquito, Zika virus, Arbovirus, Intrathoracic Inoculation, Microinjection

Experiment Summary: This protocol describes herein a protocol for the intrathoracic inoculation of ZIKV in Ae. aegypti. This method bypasses the midgut, which leads to a more rapid and higher proportion of disseminated infections in comparison to oral challenge, and mosquitoes become infected with a consistent dose of virus.

protocols
Protocol

Protocol for Delivering AAV to Express Fluorescent Biosensors in the Juvenile Mouse Hippocampus

Experiment Tags: Adeno-Associated Viruses, Acute Brain Slices, Genetically-Encoded Fluorescent Biosensors, Hippocampus, Intracranial Injection, Dentate Granule cells

Experiment Summary: In this protocol, we describe a protocol for intracranial injections of two viral vectors encoding the metabolic biosensor Peredox and the calcium biosensor RCaMP1h. When combined with 2-photon microscopy and fluorescence lifetime imaging, this protocol allows the simultaneous quantitative assessment of changes in the cytosolic NADH/NAD+ ratio and the intracellular Ca2+ levels in individual dentate granule cells from acute hippocampal slices.

protocols
Protocol

H1N1 Virus Production and Infection Protocol

Experiment Tags: Influenza A virus, Infected mouse model, H1N1 (H1N1), Virus production, Plaque assay

Experiment Summary: In this protocol, we report a protocol in infected mice with different virus doses and strains to explore how an inhibitor of lysine-specific demethylase (LSD1) impacts disease progression.

protocols
Protocol

Protocols for In vitro Transcription (IVT) and tRNA Binding Assay

Experiment Tags: T-box (T-box), EMSA (EMSA), RNA-RNA Interaction, TRNA (tRNA), In Vitro Transcription

Experiment Summary: This protocol describes the coupling of (i) “live” in vitro RNA transcription with (ii) binding by a radiolabeled, pre-formed tRNA followed by native gel electrophoresis and phosphorimager scan to visualize the complex.

protocols
Protocol

Protocol on Respiratory Syncytial Virus Infection in Mice and Detection of Viral Genomes in the Lung Using RT-qPCR

Experiment Tags: Respiratory Syncytial Virus, Respiratory Infection, Virus Detection

Experiment Summary: In this protocol, we discuss in detail how to infect BALB/c mice with RSV and how to detect the RSV genome in mouse lungs using reverse transcription quantitative PCR (RT-qPCR).

protocols
Protocol

Protocol for The Production, Titration and Imaging of Zika Virus in Mammalian Cells

Experiment Tags: Zika Virus, NS4B (NS4B), Double-Stranded RNA, Virus Titration, Immunofluorescence Microscopy, Plaque Assay

Experiment Summary: In this protocol, we describe basic experimental procedures to produce ZIKV stocks and to quantify their concentration in infectious virus particles as well as to image and study this pathogen within infected cells using confocal microscopy-based imaging.

protocols
Protocol

Chikungunya ELISA Protocol

Experiment Tags: Chikungunya, Enzyme-Linked Immunosorbent Assay (ELISA)

Experiment Summary: In this protocol, we accurately describe specific assays that can deterministically detect anti-chikungunya virus antibodies (Ab) in the tissues or sera of infected mice.

protocols
Protocol

Protocol for Transient yet Robust Expression of Proteins in the Mouse Liver via Intravenous Injection of Lipid Nanoparticle-encapsulated Nucleoside-modified mRNA

Experiment Tags: Nucleoside-Modified mRNA, Lipid Nanoparticle, Liver Regeneration, Protein Expression, Retro-orbital Injection

Experiment Summary: In this protocol, we describe in detail the necessary steps to deliver mRNA-LNP to the murine liver and, following intravenous injection of eGFP mRNA-LNP, verify transfection efficiency using flow cytometry and liver cell specificity using immunofluorescence analyses.

For research use only. Not intended for any clinical use.
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