Is there a way to improve transduction efficiency?
Is there a way to improve transduction efficiency?
Below are some tips on how to improve transduction efficiency using RetroNectin reagent.
1. We recommend preparing retrovirus supernatant at a high titer (>106 cfu/ml is desirable). To achieve transduction efficiencies of 60–70%, concentrate the retrovirus solution (e.g., measured titer = 1.15 x 106) more than 10-fold and use it for pre-loading.
2. Pre-load viral particles onto RetroNectin reagent-coated dishes at 200–250 µl/cm2, and incubate for 4–5 hours at 32°C for best results.
3. Ensure that the virus-loaded plate does not dry after discarding the supernatant and washing with PBS. Add target cells in growth medium immediately after removing the supernatant. If the plate dries, the transduction efficiency will decrease remarkably.