Muzzle warts of B6.Cg-Foxn1nu/Foxn1nu mice were
homogenized via pulverization with a mortar and pestle in liquid nitrogen then
homogenized with a tissue grinder (DWK Life Sciences, Millville, New Jersey,
catalogue no. 885450–0023). Tissue was then subjected to three
freeze-thaw cycles between liquid nitrogen and a 37˚C water bath. Tissue
was then sonicated for two minutes (amplitude = 20, 10s pulse). Cesium Chloride
(Sigma-Aldrich, St. Louis, MO, catalogue no. 289329) dissolved in
phosphate-buffered saline (PBS) was added to the wart homogenate for a final
density of 1.3623 g/mL, determined via refractometer (product discontinued).
Tissue was ultracentrifuged overnight at 36,000 rpm and opaque bands at
densities ranging from 1.27–1.31 g/mL were extracted. Extracted bands
were dialyzed three times for eight hours using Slide-A-Lyzer cassette (VWR,
catalogue no. PI66230) in three liters of PBS. The purity of the viral
preparation was confirmed using SDS-PAGE gel electrophoresis.
homogenized via pulverization with a mortar and pestle in liquid nitrogen then
homogenized with a tissue grinder (DWK Life Sciences, Millville, New Jersey,
catalogue no. 885450–0023). Tissue was then subjected to three
freeze-thaw cycles between liquid nitrogen and a 37˚C water bath. Tissue
was then sonicated for two minutes (amplitude = 20, 10s pulse). Cesium Chloride
(Sigma-Aldrich, St. Louis, MO, catalogue no. 289329) dissolved in
phosphate-buffered saline (PBS) was added to the wart homogenate for a final
density of 1.3623 g/mL, determined via refractometer (product discontinued).
Tissue was ultracentrifuged overnight at 36,000 rpm and opaque bands at
densities ranging from 1.27–1.31 g/mL were extracted. Extracted bands
were dialyzed three times for eight hours using Slide-A-Lyzer cassette (VWR,
catalogue no. PI66230) in three liters of PBS. The purity of the viral
preparation was confirmed using SDS-PAGE gel electrophoresis.