3.3. Check Toxicity of LentiTrans Transduction Reagent

The LentiTrans™ Transduction Reagent used during the transduction of target cells with lentivirus contains Polybrene, a transduction enhancement reagent. Some cell lines may be sensitive to this reagent. Before doing large-scale transduction, we recommend checking the toxicity of the LentiTrans Reagent using the procedure below:

1. In six wells of a 12-well plate, grow cells for 24 hours in complete culture medium without the LentiTrans Reagent, and with LentiTrans added at 0.25 μl/ml, 0.5 μl/ml and 1:1000 (1 μl/ml).

2. Replace old medium with LentiTrans-free complete culture medium and grow cells for an additional 72 hours.

3. Check for toxicity by counting viable cells.

For your experiments, use the highest concentration of LentiTrans Reagent that results in less than 10% cell toxicity compared to the no-LentiTrans culture (typically, a 1:1000 is recommended). For a few cell types, you may not be able to use LentiTrans Reagent.