RNAi of HepG2 cells - low transfection effeciency (Jun/21/2006 )

[size=7][font=Arial]Dear All, I was trying to perform RNAi of hGAPD (as positive control to find out the proper condition for my target gene) in HepG2 cells but failed three times. The GAPD siRNA and transfection reagent (DharmaFECT 1-DF1) were bought from Dharmacon, the cells grew well before and during the transfection, i have tried different cell density and different amount of DF1, the highest knockdown effect was just 40% (not 90% as promised by the company), i was wondering if there´s anyone who has worked on the similar project and succeeded? Any suggestion or experience would be appreciated. Help me!!!

Did you also try to increase the amount of siRNA you used? Did you check at 24, 48 and maybe 72 hours post transfection? Sometimes, using more siRNA and waiting longer for the lysis of the cells helps