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
- - - - -

RV- transfection-transduction problem

Transduction

  • Please log in to reply
No replies to this topic

#1 likepjh

likepjh

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 10 February 2015 - 04:11 PM

Hello~

 

For RV-transduction, I used calcium phosphate method.

(+ Chloroquinine during transfection/ polybrene during transduction)

Current my problem is that about 2 days after transduction, most of cells started to die.

I have no idea. Previously, it worked well, but suddenly it didn't work at all.

What I changed were HBSS and DMEM (invitro to lonza), 293T cells and I purified new plasmid DNA (endo free)

And other stocks, CaCl2, Chloroquinine, Polybrene are about 2 year old. (But keep them in -20(cacl2/chloro) and -80(poly))

 

Since non-transduced cells had better cell viability, so I think transduction may affect the cell viability.

But I have no idea what factors may affect the cell condition and viability in this method.

 

If you have any idea about this problem, please reply!!

Thanks,

 

 

 

 

 

 







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

©1999-2013 Protocol Online, All rights reserved.