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
- - - - -

Probe alignment


  • Please log in to reply
No replies to this topic

#1 kabtq9s

kabtq9s

    member

  • Active Members
  • Pip
  • 9 posts
0
Neutral

Posted 22 February 2016 - 05:52 PM

She short version:

 

This probe

WT factor V probe, HEX-ACC TGT ATT CCT CGC CT-BHQ-2

 

is not aligning to this gene sequence 

 

http://www.ncbi.nlm....531&strand=true

 

The long version

 

 

Basically we did a Taqman end point genotyping experiment to to detect Leiden Factor V Disorder.

 

We are supposed to align the primers and probe/s to the F V gene sequence.

 

Here are the primers and probes

 

Forward primer 5′-GCC TCT GGG CTA ATA GGA CTA CTT C-3′

Reverse primer5′-TTT CTG AAA GGT TAC TTC AAG GAC AA 3’

 
WT factor V probe, HEX-ACC TGT ATT CCT CGC CT-BHQ-2
 
 
This is the sequence

 

The results of the experiment indicate that we had the WT variant.

 

I am using this site to align everything

http://primer3plus.c...primer3plus.cgi

 

 

 

The primers aligned perfectly, but the probe is not aligning. I tried to reverse it and translate it but still.

 

I would appreciate any help.

Thank you


Edited by kabtq9s, 22 February 2016 - 07:55 PM.





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

©1999-2013 Protocol Online, All rights reserved.