CsCl Gradient

Use Beckman 14 x 95-mm Ultraclear tubes (Fullerton, CA, cat. no. 344060) in an SW40Ti swing-out rotor. Prepare the gradient by layering the following solutions; it is easier to put the glycerol in first, then underlay it with the p = 1.32 CsCl, underlay this with the p = 1.45 CsCl, and finally to overlay the virus. Use 2 mL of p = 1.45 CsCl, 3 mL of p = 1.32 CsCl, 2 mL of 40% glycerol, and approx 7 mL of virus prep. If necessary, fill up tube with 10 mM Tris-HCl, 1 mM EDTA, pH 8.0. Centrifuge at 80,000g for 90 min at 4°C.

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