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
- - - - -

DNA methylation Elisa kit (5-methyl-2-deoxycytidine Quantitation)


  • Please log in to reply
No replies to this topic

#1 Sarah Sobhy

Sarah Sobhy

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 18 May 2015 - 01:02 AM

Dear all,

I am master student and have DNA samples extracted from breast cancer cell lines and need to study epigenetic on them but i have quest.

 

So please check the following points that show DNA digestion step for subsequent use in methylation Elisa kit and as mentioned in my kit protocol i have to use sodium acetate and Tris on my DNA samples but unforunately did not verify the recommended volume 

 

1. Extract DNA from cell or tissue samples by a desired method or commercial DNA Extraction kit.

2. Dissolve extracted DNA in water at 0.1-1 mg/mL.

3. Convert DNA sample to single-stranded DNA by incubating the sample at 95ºC for 5 minutes and then rapidly chilling on ice.

4. Digest DNA sample to nucleosides by incubating the denatured DNA with 5-10 units of nuclease P1 for 2 hrs at 37ºC in 20 mM Sodium Acetate, pH 5.2, and following with treatment of 5-10 units of alkaline phosphatase for 1 hr at 37ºC in 100 mM Tris, pH 7.5.

 

waiting your answers.

Attached Files






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

©1999-2013 Protocol Online, All rights reserved.