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
- - - - -

use Pen./Strep. in the media for trasnfection


  • Please log in to reply
6 replies to this topic

#1 jzhen

jzhen

    member

  • Members
  • Pip
  • 3 posts
0
Neutral

Posted 08 April 2004 - 11:07 AM

Hi, all:
I am just starting to get familiar with transfection. To the media for trasnfecting the cell lines, could I add penicillian/ streptomycin in?
If not, after transfection, could I add them in? What is a good time to add P/S?
Thanks ahead
Juan
[FONT=Arial][[COLOR=purple]

#2 geness

geness

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 30 posts
1
Neutral

Posted 14 April 2004 - 06:56 AM

hi,
I am in the same situation with you!
When I transfect the cells the first time,I add P/S into the media but the transfection efficiency was very low.So the second time I do not use P/S in the media for my transfection,to my disappointment,the transfected cells were contamined mostly .

If someone know something about this,please tell us.
Thanks!

Edited by geness, 14 April 2004 - 06:58 AM.


#3 kant0008

kant0008

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 90 posts
0
Neutral

Posted 15 April 2004 - 12:03 AM

Hi guys!
I add pen/strep to all my cultures, including transfections. You're better off in most cases with a lower transfection efficiency than no cells at all due to contamination. Some transfection reagents/methods work better with some cells and not others, so I suggest use pen/strep but maybe look at other transfection methods.

#4 geness

geness

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 30 posts
1
Neutral

Posted 15 April 2004 - 03:04 AM

Thank you very much!
I agree with you!By the way,which method did you select?(chemical or physical) and I use lipofection for my transfection.I am giong to try to use CaCl2,but I wonder whether the transfection efficiency is satisfing.

#5 jzhen

jzhen

    member

  • Members
  • Pip
  • 3 posts
0
Neutral

Posted 15 April 2004 - 08:28 AM

Thank you very much!
  I agree with you!By the way,which method did you select?(chemical or physical) and I use lipofection  for my transfection.I am giong to try to use CaCl2,but I wonder whether the transfection efficiency is satisfing.

I agree too. I am using lipofectamine, which I don't know whether is the same method with lipofection. In my case, I use G418 to select the transfected cells, which has anti-G418 gene, transfected together with the gene of interest.

I was told that P/s does have some effect on the transfection. Someone suggest me that except the few hours of transfection, I should use P/S all the time.
[FONT=Arial][[COLOR=purple]

#6 kant0008

kant0008

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 90 posts
0
Neutral

Posted 15 April 2004 - 07:13 PM

Hi!
I tried Calc Phos, but it didn't seem to be particularly good. A similar methodology, but using Dextran sulfate normally works better, but there are problems with toxicity, so you got to watch it. Lipofectamine seems quite good, but you might still try smth different, like Fugene6 or DOTAP, there are tons of different reagents on the market now.

#7 fred_33

fred_33

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 288 posts
0
Neutral

Posted 15 April 2004 - 11:55 PM

Hi. For now, i've done transfection in 293 HeLa and endothelial cells. I use lipofectamine and do not add Peni/Strepto to the media for the transfection. I let the thigns happen for 3 or 5 hours (depend of the cells) then i add a complete media (ie with peni strepto) with twice the normal concentration of serum (only required if you want contihnual growth).
I never get contamination.




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

©1999-2013 Protocol Online, All rights reserved.