Selected article for: "density centrifugation and sucrose density centrifugation"

Author: Warnes, Sarah L.; Little, Zoë R.; Keevil, C. William
Title: Human Coronavirus 229E Remains Infectious on Common Touch Surface Materials
  • Document date: 2015_11_10
  • ID: 4d4l6mzl_39
    Snippet: Detection of morphological changes to HuCoV-229E using transmission electron microscopy (TEM). HuCoV-229E was purified from crude infected-cell lysate. Polyethylene glycol (PEG) precipitation (Bio-Vision PEG virus precipitation kit) was followed by sucrose density (25% to 55%) centrifugation at 96,000 ϫ g for 16 h at 4°C. The virus band was resuspended in water and the virus pelleted at 77,000 ϫ g for 1 h at 4°C. The supernatant was discarded.....
    Document: Detection of morphological changes to HuCoV-229E using transmission electron microscopy (TEM). HuCoV-229E was purified from crude infected-cell lysate. Polyethylene glycol (PEG) precipitation (Bio-Vision PEG virus precipitation kit) was followed by sucrose density (25% to 55%) centrifugation at 96,000 ϫ g for 16 h at 4°C. The virus band was resuspended in water and the virus pelleted at 77,000 ϫ g for 1 h at 4°C. The supernatant was discarded, the tubes were allowed to drain, and the final pellet was resuspended in ice-cold nuclease-free deionized distilled water after incubation on ice for 30 min. The preparation was applied to copper and stainless steel as described in the Figure 6 legend and was removed by gentle pipetting. Samples were fixed, applied to TEM grids, washed with water, and stained with 5% ammonium molybdate for 10 s.

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