Mix again and rest it on ice for 30 minutes. Add the mix precipitates into culture drop by drop, and mix gently. After 8 hrs. You should see fine precipitation under the microscope. Do not touch cells for the next 36 hrs.
Cells are weak after transfection.
On days 4 and 5 collect supernatant again. Cultured more than 3 months. Concentration of VSV-G Pseudotyped Virus Centrifuge supernatant 5 minutes at 1500 rpm. 3000 rpm x 4 C x 20' Filter the supernatant through a 0.45 um filter. Remove Supernatant by decantation and suck out residual supernatant with a pippetman.