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
- - - - -

How to remove PCR inhibitors from DNA


  • Please log in to reply
4 replies to this topic

#1 ocean

ocean

    member

  • Active Members
  • Pip
  • 24 posts
0
Neutral

Posted 25 October 2004 - 11:28 PM

hello,

does anyone know of a good way to remove pcr inhibitors during DNA extraction. currently we are using a qiagen kit and it appears that there is still inhibitors that are abolishing the pcr's i run :blink:

#2 Zaax

Zaax

    member

  • Members
  • Pip
  • 3 posts
0
Neutral

Posted 25 October 2004 - 11:48 PM

abolishing? what do you mean?

if you are getting no result, and are using the qiagen kit, i dont see what can be inhibiting the reaction.

what are you doing the pcr ON? genomic dna? (try a touchdown pcr pogram) plasmid? (try lowering the annealing temp)

make sure your primers are good.

so many things can go wrong in pcr. but if you are using kit's i dont see how inhibiters is the answer.

but i might be wrong...

#3 biomed

biomed

    member

  • Active Members
  • Pip
  • 17 posts
0
Neutral

Posted 26 October 2004 - 07:02 AM

Hi, So many things influence PCR reaction so you can not say there are PCR inhibitors in your DNA samples. Qiagen can clean the DNA or RNA sample very well. I do not think this is a issue. If you have suitable DNA control from any kit, you may try it first to explore the PCR condition for this pair of primers.
Biomed

#4 JEB

JEB

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 28 October 2004 - 04:23 AM

Hi,
Have you tried just adding some magnesium?
What you describe has happened to me simply because I was solubilizing my maxipreps in TE buffer & was presumably chelating out the magnesium needed for the PCR.
Best of luck!

#5 ocean

ocean

    member

  • Active Members
  • Pip
  • 24 posts
0
Neutral

Posted 31 October 2004 - 05:43 PM

abolishing? what do you mean?

if you are getting no result, and are using the qiagen kit, i dont see what can be inhibiting the reaction.

what are you doing the pcr ON? genomic dna? (try a touchdown pcr pogram) plasmid? (try lowering the annealing temp)

make sure your primers are good.

so many things can go wrong in pcr. but if you are using kit's i dont see how inhibiters is the answer.

but i might be wrong...

hello,

i have quantified my DNA, spectro and my 260/280 ratio is not that good (using nanodrop), 1.3-1.6. the pcr works in some run and not in others. the controls are fine, but were extracted differently. i was just wondering whether there are better kit or methods of extracting gDNA.




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

©1999-2013 Protocol Online, All rights reserved.