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!
- - - - -

should I design primers spanning both intron and exon

primer design intron exon

  • Please log in to reply
2 replies to this topic

#1 joy123



  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 105 posts

Posted 20 November 2011 - 02:38 PM

I hope to check if a gene is normal in patient. I plan to do PCR of an exon, and send for sequencing.

My question is, should I design the primers in intron, in exon, or spanning intron and exon? How should I choose among them?

Thanks a lot!

#2 pcrman



  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 1,165 posts

Posted 20 November 2011 - 06:51 PM

Do you mean you want to know if there are mutations of the gene in patients? If yes, you can just sequence exons. You can design primers on introns to span exons.

#3 Trof


    Brain on a stick

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 1,360 posts

Posted 22 November 2011 - 01:27 AM

Design the amplicon so you cover the whole exon (depending on the size, if it's too big you need to design more amplicons) plus some 50 bases of introns on each side, so you cover the whole exon in sequencing read(s). Ask for the usual read length of your sequencing facility to know how big amplicon you can have.

Our country has a serious deficiency in lighthouses. I assume the main reason is that we have no sea.

I never trust anything that can't be doubted.

'Normal' is a dryer setting. - Elizabeth Moon

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

©1999-2013 Protocol Online, All rights reserved.