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!
- - - - -

Clonetech Retro-X Tet-on system

inducible system

  • Please log in to reply
1 reply to this topic

#1 jackster101



  • Active Members
  • Pip
  • 23 posts

Posted 12 January 2015 - 04:40 PM



I'm confused! I have 2 vectors-

pRetroX-Tet-On Advanced   &

pRetroX-Tight-Pur (with my gene of interest)


My question: Tell me if I'm doing this right---I was planning on taking regular 293T cells and doing an infection into 293T (like I do for lenti)- with VSVg, deltapRetroX-Tet-On Advanced vector using Fugene--harvest the virus and infect my cells, MCF-7 and generate a G418-resistant stable line that expresses the transactivator.


I was then going to do another infection into 293T like before but use pRetroX-Tight-Pur (with my gene of interest). And then infect the MCF-7/pRetroX-Tet-On Advanced line with sup that I generated---Does that sound right?


It seems odd that I'm using the same infection protocol for retro that I have used for lenti??? 


Are infection protocols similar. I remember from years ago--people would use Phoenix cells than 293T

#2 Robin Shen

Robin Shen


  • Members
  • Pip
  • 2 posts

Posted 01 April 2015 - 01:55 AM

The first process is correct. 

First is select stable clone which stable express rtTA, and next step is transfect CMVtet-Target gene to rtTA stable clone.

Would be a waste of time. 

You also can construct double promoter vector which express rtTA and target gene at same time.

Also tagged with one or more of these keywords: inducible system

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

©1999-2013 Protocol Online, All rights reserved.