WinterVomitingLab:Protocols

WinterVomitingLab

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General Virus Methods

Virus Concentration

• PEG precipitation
• Amicon ultrafiltration
• Ultracentrifugation

Virus purification

• Organic solvent extraction

Virus stock preparation

• From a stool or vomit sample
• Partially purified cell culture lysate

Cell Culture

Cell passage

• RAW 264.7 cells
• CRFK cells
• FRhK-4 cells
• MDCK.2 cells
• HeLa cells
• A549 cells

Propagation (making dishes)

• For MNV-1
• For FCV or HAV

Virus Culture

Flask infection

• MNV, FCV or HAV flask infection

Plaque assays

• MNV-1 plaque assay
• FCV plaque assay
• HAV plaque assay

Cell/plaque staining

• Crystal violet
• Neutral red

Viral Nucleic Acid Methods

Extraction methods

• Heat release

Histo-blood group antigen binding assays

• Using NeutrAvidin plates
• Using magnetic beads

Disinfection/Sanitizing

• Virus removal from berries using sanitizers
• In-solution inactivation test