Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
Photo

Protocol for HT29 siRNA transfection using Lipofectamine 2000?

HT29 siRNA Lipofectamine 2000 transfection

  • Please log in to reply
3 replies to this topic

#1 ammahew

ammahew

    member

  • Active Members
  • Pip
  • 10 posts
0
Neutral

Posted 19 September 2012 - 02:19 AM

Hi everyone,

Can anyone suggest a protocol for transfection of HT29 cells with siRNA using Lipofectamine 2000?

I have successfully transfected HCT116 cells, but have been told HT29s are more difficult to transfect. I'll try using the same protocol as the HCT116s initially, but I was wondering if anyone had any experience transfecting HT29s - are they more difficult to transfect? Will I need to increase the siRNA/Lipofectamine concentration, or transfect for longer?


Thanks!

#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,733 posts
400
Excellent

Posted 19 September 2012 - 02:03 PM

You will need to optimize the transfection for your purposes; each cell line is different. I would recommend however, that you get some lipofectamine RNAiMax instead of 2000, it is a lot less toxic and seems to work really well for siRNA transfection.

#3 ammahew

ammahew

    member

  • Active Members
  • Pip
  • 10 posts
0
Neutral

Posted 21 September 2012 - 01:16 AM

Ok, I think I might actually have some RNAiMax somewhere from an old lab, thanks!

#4 vitalgene

vitalgene

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 53 posts
0
Neutral

Posted 26 September 2012 - 12:05 PM

i would suggest a reverse trasfection for HT29 cells.





Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.