I have generated the lentivirus particles with my GOI. But did not get transduction efficiency in Jurkat cells post 72 hrs why? any suggestion ?
Particulas: used 24 well plate :
50,000 cells with Polybrene 8ug/ml
Thank you in anticipation
Yes, functional titration done.
lentivirus generated in 60 mm dish, concentrated (PEGit), finally dissolved in 50ul PBS.
Transduced Jurkat cells and HEK 293 cells with 5ul virus each. Observed 90%GFP in HEK293 (50,000 cells-by this I would say 2*10^5 TU/ml titer got) but not in Jurkat cells (even after spinoculation), why so?
In conclusion,
(50,000 cells >5ul virus> 1 lakh TU= 2MOI in Jurkat cells)
Dear Tom Masi,
Kindly let me know how you worked on transduction in Jurkat cells.
what are the factors that i should take care on?
Thank you very much in anticipation....
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