Jump to content

  • Log in with Facebook Log in with Twitter Log In with Google      Sign In   
  • Create Account

- - - - -

Ammonium acetate DNA extraction mistake

DNA extraction

  • Please log in to reply
1 reply to this topic

#1 Click

Click

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 27 October 2011 - 06:50 AM

Hi all,

I am using an ammonium acetate extraction protocol to extract DNA from some insect samples. However, instead of adding Digsol to the chopped up sample, I made a mistake and added ammonium acetate (this should have been done in the step after digestion!). I panicked and added the digsol anyway, so now I have a tube containing the sample, digsol, ammonioum acetate and proteinase K currently incubating at 55oC.

Does anyone know whether I've messed up my extraction by doing this and if so is there a way of rescuing it? Or can I carry on with the protocol as normal (i.e. leave to digest overnight, then add more ammonium acetate, then centrifuge and carry out the ethanol washing steps)?

Any advice would be appreciated!

Thanks

#2 hobglobin

hobglobin

    Growing old is mandatory, growing up is optional...

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 5,252 posts
60
Excellent

Posted 27 October 2011 - 07:51 AM

No idea if this works or not, but Proteinase K is quite stable and should survive this. At least I'd check the pH and adjust it to the pH of the extraction buffer to avoid a too low value.
  
One must presume that long and short arguments contribute to the same end. - Epicurus
...except casandra's that belong to the funniest, most interesting and imaginative (or over-imaginative?) ones, I suppose.





Also tagged with one or more of these keywords: DNA extraction

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

©1999-2012 Protocol Online, All rights reserved.