I am trying to express GOI in Min6 (mouse) cell line using Chloroquine and PEI (1:3) protocol from https://www.addgene....rus-production/.
I have tried using 293T, LX293T and even 293FT cells for virus production.
I have collected the virus every 24 hrs (like the Addgene protocol says) and also leaving them for 72 hrs straight and then collecting.
I filter the supernatant using 0.45 um syringe filter and use Polybrene (0.8 ug/ml) for transduction.
I have also tried "spinfection" after reading some answers on this forum.
My problem is I see a lot of GFP upon transfection indicating good transfection efficiency. After collecting the viral supernatant when I transduce Min6 cells I see little to no GFP in them. Where am I going wrong?