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
* * * * * 1 votes

Amount of positive control for bisulfite treatment


  • Please log in to reply
1 reply to this topic

#1 methdetector

methdetector

    member

  • Active Members
  • Pip
  • 11 posts
0
Neutral

Posted 11 December 2009 - 01:40 PM

Hello everybody,

I am using EZ DNA Methylation-Gold Kit for bisulfite treatment of plant DNA. I plan to use 'Universal Methylated DNA Standard and control primers' from Zymo research in conjunction with it as a positive control. I understand that a minimum of 500 pg of DNA is required for bisulfite conversion. However, the concentration of Universal Methylated DNA Standard indicated on the tube containing the DNA provided is 100pg/20uL (5pg/uL). But it is also mentioned that there is 5ug/20uL of salmon sperm DNA as a carrier. I am confused as to how much of standard control DNA would be required for treatment. :D

Any suggestion would be greatly appreciated. Hope to hear from somebody soon :D

#2 pcrman

pcrman

    Epigenetist

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 1,165 posts
68
Excellent

Posted 13 December 2009 - 12:49 AM

So the concentration (5 pg/ul) is the already modified DNA. If you want to know how much DNA should be used for treatment, ideally you can use about 1 ug DNA for treatment. Because there is significant DNA loss during treatment and purification, the recovered DNA from 1 ug starting DNA after treatment would be at picogram scale. I guess that is why the standard modified DNA provided in the kit has such low concentration.




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

©1999-2013 Protocol Online, All rights reserved.