1 reply to this topic

[synbionoob]

Hey everyone,

I am trying to transfect ER+ mouse breast cancer cellls using DharmaFECT transfection reagent and Dharmacon siRNA, but dont seem to be getting any more than 30% inhibition. Any ideas on how I can improve the efficiency?

Thanks!

[Dr. Zhang]

synbionoob, on 08 March 2011 - 07:19 PM, said:

Hey everyone,

I am trying to transfect ER+ mouse breast cancer cellls using DharmaFECT transfection reagent and Dharmacon siRNA, but dont seem to be getting any more than 30% inhibition. Any ideas on how I can improve the efficiency?

Thanks!

Just some tips:
1)cell status and confluency
2)appropriate siRNA conc.(the newest Dharmacon siRNA transfection protocol recommonds a final conc. of 25nmol)
3)resuspend the siRNA powder with RNase free water(DEPC water or siRNA resuspension buffer supplied by Dharmacon just ok)
4)always use filter-tips to gently handle siRNA

Just go back to the protocol of Dharmacon siRNA and follow them strictly, you should easily get a nearly 100% mRNA konckdown efficiency at 48~72hrs post transfection.