WinterVomitingLab:Protocols

WinterVomitingLab

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General Virus Methods

Virus Concentration

• PEG precipitation
• Amicon ultrafiltration

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

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