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Prepare transfection reagent A: add 1ul Lipofectamine into 30ul serum-free culture medium
Reagent B: add Tug expression plasmid + 25ul SIRNA into 30ul serum-free culture medium
Mix reagent A and B
Before transfect, replace with fresh serum-free medium
Add mixed transfection reagent
Incubate at 37 C for 4 hours
Wash cells by 500ul 1X PBS
Add 70ul 1x sample buffer
Harvest cells
Incubate at 90°C for 10 minutes