No replies to this topic

[kabtq9s]

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

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

 

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.