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
- - - - -

qPCR - no amplification curve but suitable melting curve


  • Please log in to reply
3 replies to this topic

#1 pcr_martin

pcr_martin

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 17 July 2013 - 11:59 PM

Hi there,

last week i run a qpcr-probe (two step, 40 cycles); there was no amplification , then i analysed the melting curve an there was a really suitable peak!

what´s going wrong there? I would be really grateful to have suggestions from you!

greets,
Martin

#2 Podge

Podge

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 47 posts
2
Neutral

Posted 18 July 2013 - 05:46 AM

Hi there,

last week i run a qpcr-probe (two step, 40 cycles); there was no amplification , then i analysed the melting curve an there was a really suitable peak!

what´s going wrong there? I would be really grateful to have suggestions from you!

greets,
Martin


Do you still have the plate? You could put the reaction on to an agarose gel and see if there is an amplicon as indicated by the melting curve.

#3 pcr_martin

pcr_martin

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 18 July 2013 - 11:05 PM

Hi,

I´ve done it, there was a amplicon as indicated; attached you´ll find a scrrenshot; the red peak is the first band, the green one the second band

Attached Thumbnails

  • 1 ng.JPG
  • 1ng amp.JPG
  • DSC_4938a.JPG

Edited by pcr_martin, 18 July 2013 - 11:09 PM.


#4 biothreat

biothreat

    member

  • Members
  • Pip
  • 4 posts
0
Neutral

Posted 20 September 2013 - 04:30 PM

It seems as though your probe is not binding.






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

©1999-2013 Protocol Online, All rights reserved.