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
- - - - -

solubility of dna


  • Please log in to reply
4 replies to this topic

#1 samm

samm

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 27 June 2004 - 04:10 AM

help!
im looking for the best method to dissolve dna-pellets,after it is extracted. the pellet is not completely dissolved in TE ph 8,0 and therefor the pcr shows not the results it should be. :( do you the best way to dissolve the dna-pellet??please tell me!!thank u..

#2 hula

hula

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 30 posts
1
Neutral

Posted 27 June 2004 - 10:14 AM

What method did you use for your DNA extraction? Are you sure the unsoluable pallet is DNA? What does it look like?

#3 kant0008

kant0008

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 90 posts
0
Neutral

Posted 27 June 2004 - 04:35 PM

Hi!
I agree with hula, it could be something other than dna that you've isolated. Otherwise could be a number of things:
1) Did you have an ethanol wash as the last step of your protocol? If the ethanol wasn't dried off properly your nucleic acid will not dissolve. I normally dry my samples with a pipete tip (capillary action, upside down) and then leave them for a while with lids open (cover with tissue or something) on bench or ona heating block (not too hot). With the samples you alredy have you can do an ethanol precipitation and redissolve
2) Your dna might be very concentrated and so reached the extent that it'll dissolve, what's the concentration? Try adding more buffer.
3) Leave your dna at 4C overnight to allow to dissolve, genomic dna takes a while to dissolve
Hope this helps!

#4 hula

hula

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 30 posts
1
Neutral

Posted 27 June 2004 - 04:56 PM

also if your DNA is high molecular DNA and looks viscid, dissolve it in 8mM NaOH solution, or 20 to 500ul to 10mM Tris HCl, 0.1mM EDTA, pH 7.5. Complete dissolution of the DNA may requires several hours of agitation at 37C or 55C (may need overnight).

#5 Calamhex

Calamhex

    member

  • Members
  • Pip
  • 3 posts
0
Neutral

Posted 29 June 2004 - 05:49 PM

If an overnight incubation in TE does not dissolve the stuff that you see... my best advice is to remove that stuff, which is probably just junk by centrifuging and transfering clear supernatant to a tube and then retry your PCR. The junk may contain PCR inhibitors...
Also, sometime with DNA dissolved in TE one has to slightly increase MgCl2 concentration... usually 0.25 mM is enough... depends on volume of DNA you are adding though...

HTH




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

©1999-2013 Protocol Online, All rights reserved.