Lentivirus Purification Ultrafiltration

1. After PEG/NaCl precipitation and centrifugation, a white pellet should be visible.

2. Carefully decant the supernatant and add 1ml 5 ml PBS (or other buffer) to the centrifuge tubes.

3. Resuspend the pellets by pipetting liquid gently up and down.

4. Transfer suspension to 100KDa spin concentrator.

5. Add 2 X 5 ml of PBS to wash centrifuge tubes, transfer liquid to 100kDa spin concentrator.

6. Spin at 3000 rpm for 15 min to 1 hour, in cold room

7. Add 10 ml PBS to spin concentrator, spin for another 15 min to 1 hour.

8. Transfer remaining liquid into screw-cap microfuge tubes in aliquots of 200 ul.

9. Add 1 ml PBS to wash the spin concentrator. Do not aliquot this wash.

10. Titer virus by RT-PCR, or 293 cell infection or p24 ELISA.

11. Print labels (Catalog number, Product/virus name, Lot #, Date, Titer, Company Name).

12. Snap-freeze the tubes in crushed dry ice and store at -80 oC.