Decoration Using Crude Sap Extracts

1. Crush a small amount of virus infected material (see Note 9) in 1020 pL of distilled water on a glass slide.

2. Touch a carbon-coated grid (see Note 6) on to the preparation and remove after a few seconds.

3. Wash with 30 consecutive drops of phosphate buffer pH 6.5 (see Note 11), draining with filter paper but do not dry.

4. Dilute specific antiserum to 1:100 (see Note 3) in a 0.5-mL microfuge tube and place 20-pL drops on Parafilm in a Petri dish.

5. Wash the grid with 20 drops of buffer as before and drain as before. Place it on the diluted antibody and incubate for 15 min at room temperature (see Note 7).

6. Wash the grid with 20 drops of distilled water and stain with three to five drops of 2% uranyl acetate (see Note 12), draining as before (see Note 14).

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