Traditionally the recombinant lentivirus is purified by ultracentrifugation to separate the virus particles from cellular proteins and media components. The ultracentrifugation procedure is time consuming and limited to the amount of cell lysate to be processed. The Lentivirus Concentration Reagent is designed for fast and efficient concentration of recombinant lentiviruses from lentiviral-transfected cell culture supernatant. The virus can be concentrated 50 -100 folds. The recovery rate is around 60-70%.
Storage and Stability
The guaranteed shelf life is 12 months from the date of purchase. Store all components at room temperature (15-25℃).
Kit Components
Catalog#
BMG-V2001-00
BMG-V2001-01
BMG-V2001-02
Buffer LP
10 mL
100 mL
500 mL
Buffer LS
1 mL
10 mL
50 mL
Protocol
Centrifuge the lentivirus-infected culture media at 3,000 rpm for 10 minutes at 4℃. Filter the supernatant through a 0.45 μm filter. Supernatant from 1-2 T75, up to 30 mL of supernatant, can be processed per prep. Note: The supernatant can also be stored at -80°C for future purification.
Add 1 volume of Buffer LP to 3 volume of virus supernatant (For example, add 5 mL of Buffer LP to 15 mL of virus supernatant). Mix well and incubate at 4°C for at least 4 hours to overnight. The virus is stable in Buffer LP.
Centrifuge the sample at 3,000 rpm for 30 minutes at 4°C. Carefully aspirate the supernatant. Spin briefly and remove the residual supernatant. The virus containing pellet should be visible. The pellet may appear hazy. Keep the virus on ice and proceed to step 4.
Resuspend the pellet with 300–500 µL Buffer LS. Dissolve the pellet by pipetting. Transfer the pellet to a clean vial and spin at 8,000 rpm for 2 min at 4°C.
Transfer the supernatant to a clean vial. Aliquot and store the purified virus at -80°C.
