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A simplified purification method for AAV variant by polyethylene glycol aqueous two-phase partitioning | Cell Pellet→Lysis→Nuclease→PEG8000-NaCl Precipitation. Replace from ultracentrifugation to precipitation procedure | 10% PEG8000, 13.2% (HN4)2SO4 | |
Culture Supernatant + PEG solution (1/4 vol)→stir slowly 4℃, 1hr→4℃, 3hrs without string to allow full peciptation | 40% PEG 8000, 2.4% NaCl, adjust pH7.4 | ||
Adeno-associated Virus (AAV) AssemblyActivating Protein Is Not an Essential Requirement for Capsid Assembly of AAV Serotypes 4, 5, and 11 | make in 8% PEG 8000, 0.5 NaCl on ice for 3hrs. The pelet was suspet by 50 mM Tris-HCl (pH 8.5) and 2 mM MgCl 2 | ||
Helper-free Production of Laboratory Grade AAV and Purification by Iodixanol Density Gradient Centrifugation | add 1:4 vol precipitation solution to supernatant from cell culture | 40% PEG 8000, 2.5 M NaCl water |