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
- - - - -

ABI377 Sequencing of PCR products


  • Please log in to reply
1 reply to this topic

#1 deano

deano

    member

  • Members
  • Pip
  • 4 posts
0
Neutral

Posted 07 June 2002 - 09:52 AM

I have great success manually sequencing PCR products with the USB Terminator Cycle Sequencing kit. I would like to switch over in some cases to using the ABI377 automated sequencer at the center on campus for some of these products, but I have had very poor results with it. The signal strengths are weak and the calls are difficult to make with high background. Does anyone sequence PCR products on a regular basis that can give me some suggestions for improved results? I normally sequence my products with the same primers used for amplification - I have not tried nesting sequencing primers. I have also used the TOPO TA kit and have had mixed results with sequencing those products as well. All the products are cleaned up with either Qiagen or Bio-Rad kits, so I don't think contamination is a problem.

Little help?!?


#2 deano

deano

    member

  • Members
  • Pip
  • 4 posts
0
Neutral

Posted 14 June 2002 - 06:21 AM

I must be the only one who reads this board who sequences PCR products. Thank you for all your help everyone!




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

©1999-2013 Protocol Online, All rights reserved.