AAV sample does not flow through the column due to one or more of the following:
Centrifugation speed was too low. Check the centrifuge to ensure that it is capable of generating 1,500 x g. Sufficient centrifugal force is required to move the liquid phase through the resin.
Inadequate spin time. Spin an additional two minutes to ensure that the liquid is able to flow completely through the column.
Cell debris obstructing column. Ensure that no cell debris is inadvertently applied to the column following lysate centrifugation. Care should be taken to ensure that only the supernatant is applied to the column. Filtration of the supernatant through a 0.2 µm or 0.45 µm filter may decrease clogging of the column. The sample can also be divided among several columns.