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
- - - - -

Transcription of PCR products without a primer coupled with promotor sequence


  • Please log in to reply
4 replies to this topic

#1 GradMicro

GradMicro

    member

  • Active Members
  • Pip
  • 24 posts
0
Neutral

Posted 02 December 2012 - 07:03 PM

Hello,

Is there a way to transcribe PCR products wihtout using a primer that has a transcription promotor. I plan to do base specific cleavage followed by MALDI TOF for strain differentiation. My PCR is a multiplex PCR with 15 primer pairs. Thus, tagging the primers is not a possibility.
Any alternatives are most welcome.

Thank you

#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,236 posts
336
Excellent

Posted 03 December 2012 - 11:47 PM

Why do you need to transcribe? surely this is essentially the same as doing a PCR with Uracil.

#3 GradMicro

GradMicro

    member

  • Active Members
  • Pip
  • 24 posts
0
Neutral

Posted 04 December 2012 - 05:40 AM

All protocols for MALDI TOF based sequencing using base soecific cleavage is by this method. The cleavage is by RNAses, I think that is the reason.

#4 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,236 posts
336
Excellent

Posted 04 December 2012 - 11:24 AM

Ah, fair enough. You would definitely need a promoter to get this to work. Would it be possible to clone (possibly topo-TA) the PCR products?

#5 GradMicro

GradMicro

    member

  • Active Members
  • Pip
  • 24 posts
0
Neutral

Posted 04 December 2012 - 01:00 PM

That is possible. However, if there are quicker methods, it is better as the final objective would be to see if the method can be diagnostically applicable. As cloning would take additional days is not the best option.




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

©1999-2013 Protocol Online, All rights reserved.