Hiperfect siRNA in HT-29 cells help - (Oct/16/2012 )
Does anyone have experience in using hiperfect transfection reagent in HT-29 cells?
So for I've used 5, 10, and 25mM siRNA with 3, 4.5 and 11ul Hiperfect for 48hrs, but am not getting any knockdown. I'm seeding on 24 well plates at 180,000 cells/well.
I was going to start again with 35mM and 50mM and 9ul hiperfect at 48h and 72hrs. But if anyone has any tips or ideas that would be great, as it would save me precious time!
Many thanks for any info
Are you working with a siRNA that is known to cause knockdown? If not, it may not be a problem with your transfection, and you should get a known working control.
It works really well in DLD cells - knocks the gene down by ~96%. There are about 4 times as many HT29 cells in one well than DLDs though so I wondered if volume had something to do with it too