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
- - - - -

Fugene transfections


  • Please log in to reply
1 reply to this topic

#1 Suzanna

Suzanna

    member

  • Active Members
  • Pip
  • 10 posts
0
Neutral

Posted 11 October 2012 - 09:54 AM

I think I must be really stupid but my effeciencies with calcium phosphate actually work better than fugene/lipofectamine!! What is a typical protocol for fugene transfection ? does adding fugene to the diluent with dna make a difference than making separate dilutions of dna and fugene in serum free medium and mixing ? also what is the typical complexing time ? what efficiency should i expect for an endothelial cell line (p9)? any tips and suggestions welcome

thanks

Suzanna

#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,784 posts
406
Excellent

Posted 12 October 2012 - 07:38 PM

Fugene in the presence of water forms small charged droplets called lipospheres with the DNA inside them due to charges on the DNA that are not present in the medium/water (medium is mostly water). You should be adding the fugene directly to the DNA/diluent mix.

Complexing should be 15-45 min, but can be longer.

Can't comment on the endothelial line though.




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

©1999-2013 Protocol Online, All rights reserved.