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[EXTERNAL] [Zbrafish] Nacre primers

James Lister via zbrafish%40net.bio.net (by james.lister from vcuhealth.org)
Thu Feb 8 13:53:52 EST 2018

Hi again Cameron,

Also, the 19bp deletion in roy is actually in the mpv17 transcript (due to mis-splicing) not in the genomic DNA.  To my knowledge the mutation itself has not been identified, so you’ll have to use non-complementation or sequencing from cDNA to ID those carriers.


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>

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