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
- - - - -

Reducing DNA sample viscosity of TCP

DNA Lysis Total Cell Pellet

  • Please log in to reply
No replies to this topic

#1 Missle

Missle

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 132 posts
18
Good

Posted 31 July 2013 - 07:31 AM

Hello.  I am looking to analyze the expression of my bacterially expressed recombinant protein in the total cell pellet (TCP) fraction in a plate format.  I've collected my bacterial pellet within the pellet, resuspended, added SDS-PAGE sample buffer and boiled.  The problem is that my resultant TCP fraction is too viscous to pipet.  Sonication is not an option because of my sample number and lack of high throughput sonication equipment.  I used a syringe and needle, which did help, but this is not a long-term solution as I am processing 30+ samples and will consume lots of syringes and needles and it will take a long time (kindof puts a kink in the high throughput part of my analysis :(

 

In the past I've used DNase to solve this problem during lysis but am assuming that DNase won't function in the presence of SDS.

 

Does anyone have any suggestions of how to reduce my viscosity while retaining my throughput?

 

Thanks!!!







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

©1999-2013 Protocol Online, All rights reserved.