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
- - - - -

PCR blood old samples


  • Please log in to reply
2 replies to this topic

#1 Aline

Aline

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 28 November 2004 - 10:46 AM

Hi everyone,
Please, I have a problem and I need help! I have several blood samples obtained since 2002. These samples were mantained at 4ēC for several months, and at -20 ēC since may 2004. I´beem trying to extract DNA from these samples with DNAzol and the amount obtained was about 0.5 microgram/microliter from 100 microliter of blood. the samples were collected in EDTA tubes.
My question is: it is possible to perform PCR with these samples? The time of storage could degrade the DNA? the quantification in spetrophotometer could show contamination with proteins or other compounds masquerading the amount of DNA quantificated?
Thanks a lot for your attention!


Aline

#2 nabla

nabla

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 41 posts
0
Neutral

Posted 28 November 2004 - 11:00 PM

Hello! The problem is less the freezing but the storage at 4 °C. But I do not think ist is absolutely unrealistic, just try. You will probably will be surprised about the good results. But you have to be shure that all EDTA is removed. Further more it depends on the amplicon size you want to get, about 200 bp should be no problem. Of course the DNA will be degraded. Have you determinded the ration A 260 nm to A 280 nm? Than you will get more Information about possible contamination with organic solutions (phenol) or proteins.

Edited by nabla, 28 November 2004 - 11:00 PM.


#3 mabacil

mabacil

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 01 December 2004 - 08:32 AM

try to run your isolated DNA on 1 or 2% agarose gel. if you will see one band, your DNA is not degraded. if you will see ladder it is.. but even on degraded DNA is possible to run PCR.. just try it. good luck!




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

©1999-2013 Protocol Online, All rights reserved.