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
- - - - -

Direct sequencing of Bisulphite PCR product


  • Please log in to reply
1 reply to this topic

#1 ameybendre

ameybendre

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 25 February 2010 - 07:06 PM

HI,
i have done bisulphite treatment and have got excellent MSP bands.i want to proceed bisulphite sequencing using BSP's.is there any specific protocol or steps when u have to perform direct sequencing ?

#2 pcrman

pcrman

    Epigenetist

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 1,165 posts
67
Excellent

Posted 25 February 2010 - 09:42 PM

You will need different primers from MSP primers to amplify the modified DNA. Once you get PCR products, send them for sequencing. To get enough products for sequencing, most likely you have to do nested or two rounds of PCR. You can use either forward and reverse primer for sequencing, but reverse primer always works better than forward one.




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

©1999-2013 Protocol Online, All rights reserved.