Lentiviral production

Starting from 5 x 1ml/vial of 293T serum-free supernatant

Tittered 3E+7 to 1E+8IU/ml without centrifuge/filter concentration

Diagram showing a process involving labels, a petri dish with cells, and particles moving toward the dish.

Biosettia have developed a unique way to determine the functional titers.

Transduce 5E+5 H1299 cells/well in 6-well plate with 20 ul of lentiviruses in 2 ml DMEM + 10% FBS and 8 ug/ml polybrene, spin at 1,000 g for 60 min, extract genomic DNA around 16 hr post-transduction for real-time PCR analysis. The GFP-Bsd lentivirus pre-titered in H1299 cells by serial dilution and Blasticidin selection is used as reference copy number to determine the functional titers of lentivirus samples. The primers targeting BRCA1 locus is used to determine genomic DNA copy number for normalization. Biosettia’s virus titering method determines the copy numbers of reverse transcribed lentiviral genome inside of the transduced H1299 cells which is more accurate than p24 ELISA and viral RNA qPCR, and time saving than antibiotic selection from virus serial dilution.