Editing Adenovirus Purification Clontech (section)

== Amplify Adenovirus in HEK 293 Cells<br>  ==

1. Detach low passage HEK 293 cells with trypsin, wash, and count. 

2. In a 50 ml conical tube, dilute the cells to a final density of 1.83x10^6 cells/ml in a total of 40 ml. 

3. Add virus to the cells and mix. 

Note: Perform the test titer in Part A to determine the optimal amount of virus to add in this step. 

4. Aliquot 33 ml of fresh medium onto each of five 150 mm cell culture dishes. 

5 Aliquot 8 ml of virus-infected cells (from step 3) onto each of the five 150 mm cell culture dishes; make sure the cells are evenly distributed.<br>Note: This is equivalent to 1.46x10^7 cells/plate. 

Incubate the cells at 37oC and 5% CO2 until the CPE is complete; this should take approximately 3-5 days. 

Note: Optimally, the purification assay should be performed when 50% of the cells are detached, and the remainder are attached but rounded.