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
- - - - -

Multiple missense mutations in a single exon?

Sanger Sequencing

  • Please log in to reply
3 replies to this topic

#1 Ameya P

Ameya P

    Rervm Cognoscere Cavsas

  • Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 330 posts
25
Excellent

Posted 14 March 2014 - 06:59 AM

Just wanted to know if any one one had come across multiple missense mutations in a single exon for a single sample. 

 

Just got back my Sanger Sequencing results and one of the reads has like 3 missense mutations in a single read (both directions). These do not look like some artifact and have been very clearly called. So, just wanted to know what are the chances that something like  this might happen. 


NEW!!!!  5 Reasons why Rosetta is a SUPERSTAR on CoffeeTableScience!!!!

Image copyright: Adrian Koh SF.
Replication of this art is strictly prohibited without express permission of the artist


#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,597 posts
384
Excellent

Posted 14 March 2014 - 10:28 PM

Is this from cloning or what?



#3 Ameya P

Ameya P

    Rervm Cognoscere Cavsas

  • Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 330 posts
25
Excellent

Posted 15 March 2014 - 01:19 AM

No, PCR product. Purified prior to sequencing.  But homozygous missense mutations observed in the middle of the read, in both directions


NEW!!!!  5 Reasons why Rosetta is a SUPERSTAR on CoffeeTableScience!!!!

Image copyright: Adrian Koh SF.
Replication of this art is strictly prohibited without express permission of the artist


#4 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,597 posts
384
Excellent

Posted 16 March 2014 - 11:51 AM

In that case, no, I haven't come across it. I have seen things like this with cloning, where the proper product is toxic, but somehow the bacteria have mutated the insert such that it no longer is toxic.






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

©1999-2013 Protocol Online, All rights reserved.