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
- - - - -

Rescue my DNA extraction


  • Please log in to reply
1 reply to this topic

#1 mray

mray

    member

  • Active Members
  • Pip
  • 10 posts
0
Neutral

Posted 15 October 2009 - 12:19 PM

Hi,

I was trying to carry out DNA extraction, but I screwed up while EtOH ppting. I added 70% EtOH (for rinsing), and vortexed by mistake!! So my DNA went back into solution. I centrifuged it again (max rpm, 15mins) and pellet was much smaller (I guess because pption is not efficient from 70% EtOH, and I didn't have any salt). Is there any way to fix this??

I was considering adding 100% EtOH till conc was 95% at least, then adding Na-acetate and incubate @ -20C before spinning. But then the total vol becomes large - 5mL and DNA conc will be very low, but maybe if I add glycogen I can still recover some DNA? Also how to I calc how much salt to add. Normally I add 1/10th of vol of DNA solution, but now my solution is in 70% EtOH not buffer...

Please help!! My sample is very precious since its from a tumor.

Thanks!

#2 hanming86

hanming86

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 156 posts
2
Neutral

Posted 15 October 2009 - 09:09 PM

u are supposed to etoh wash + vortex. so that the pellet is resuspended and the contaminant is all washed off. then the centrifuge step to get the pellet back again.
Lab + Coffee + Music = Bliss




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

©1999-2013 Protocol Online, All rights reserved.