High Efficiency Lenti2go

Lentiviral Particle Assembly & Infection Kit

 

The High Efficiency Lenti2go Lentiviral Particle Assembly and Infection Kit provides a convenient and efficient tool for producing lentiviral particles to deliver and express your gene of interest in dividing, non-dividing, or primary cells. The multi-vector pre-mixed Packaging Mix is easy to use and designed to express viral structural proteins and the VSV-G (vesicular stomatitis virus glycoprotein) envelope protein at ratios optimal for lentiviral assembly and production. In addition, the Kit also provides a lentiviral transduction enhancer, Lenti-InfectinTM, a proprietary technology developed at Virongy, to significantly increase productive viral transduction rate (up to 30-fold). The use of Lenti-InfectinTM eliminates the need to concentrate viral particles, and greatly facilitates biological and biochemical characterization of lentiviral transduced cells.

 

Highlights

  • Preoptimized vector mix, ready to use, and easy steps
  • Lenti-InfectinTM technology that enhances transduction efficiency up to 30-fold
  • No need for viral particle concentration
  • High transduction efficiency for both cell lines and primary cells.

 

 

 

Protocol

Lentiviral particle assembly using Lenti2go Lentiviral Kit (10 cm dish)

  1. One day (18-24 hours) prior to co-transfection, plate 3 x 106 HEK293T cells in a 10 cm dish and grow at 37°C to reach about 80% confluence.
  2. Remove medium from the dish and rinse with serum free DMEM medium, then add 9 ml of warm, serum-free medium.
  3. Prepare transfection solution: Set up two tubes (A) and (B). (A) is for DNA dilution, and (B) is for dilution of Transfectin.
  4. In (A), add 5 μg of Packaging vector Mix, add 5 μg of Lent-viral vector of your choice, and then add serum-free DMEM medium to make a final volume of 500 μl, Mix well.
  5. In (B), add 45 μl of Transfectin, and then add 455 μl if serum-free DMEM medium to make a final volume of 500 μl. Mix well.
  6. Combine (A) and (B) into a total volume of 1 ml. Mix DNA and Transfectin by vortexing, and incubate at room temperature for 15 min.
  7. Add the 1 ml (A) + (B) mixture to the dish drop-wise. Mix gently by slowly rotating the dish. Incubate at 37°C for 6 hours.
  8. Carefully remove the supernatant with a pipette (careful not to remove cells). Add 10 ml of warm DMEM with serum. Culture cells for 48 hours.
  9. Harvest virus at 48 hours. Filter through a 0.45 μm to remove cellular debris. Aliquot and store the virus at -80°C.