IUBio Biosequences .. Software .. Molbio soft .. Network News .. FTP

[EXTERNAL] [Zbrafish] Nacre primers

James Lister via zbrafish%40net.bio.net (by james.lister from vcuhealth.org)
Thu Feb 8 13:28:01 EST 2018

Hi Cameron,

Actually w2 is a substitution that fortuitously creates a Dra I restriction site, so the genotyping is pretty straightforward and you can use pretty much any primers to your liking that flank the mutation (although be aware there is another Dra I site not too far away.)  I can send you our primer info/protocol if you like.


On Feb 7, 2018, at 1:11 PM, Cameron Wyatt <Cameron.Wyatt from igmm.ed.ac.uk<mailto:Cameron.Wyatt from igmm.ed.ac.uk>> wrote:

Dear all,

Does anyone have experience genotyping nacre mutants (mitfa^w2) by PCR?

I have pigmented offspring of casper hets that I would like to genotype. Designing primers for the roy mutation (mpv17^a9) appears to be easy enough as it is a 19bp deletion. However the nacre mutation is a 1bp substituition. I've never used PCR to identify a 1bp mutation and I've read that 'sometimes it works, sometimes it does not'.
Does anyone know if PCR works for this particular mutation (presumably with a primer 3' aligned with mutant bp)?

To save people the time of writing to suggest alternatives, I will say that I know can sequence the region containing the mutation or pair mate the fish with homo caspers instead.

Thanks for any advice.


Cameron Wyatt
Zebrafish technologist and manager
HGU fish facility
Institute of Genetics and Molecular Medicine
University of Edinburgh
The University of Edinburgh is a charitable body, registered in
Scotland, with registration number SC005336.
Zbrafish mailing list
Zbrafish from net.bio.net<mailto:Zbrafish from net.bio.net>

James Lister, Ph.D.
Department of Human and Molecular Genetics
Virginia Commonwealth University School of Medicine
Richmond, VA 23298 USA
James.Lister from vcuhealth.org<mailto:James.Lister from vcuhealth.org>

-------------- next part --------------
An HTML attachment was scrubbed...
URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20180208/ae5a1efd/attachment-0001.html

More information about the Zbrafish mailing list

Send comments to us at biosci-help [At] net.bio.net