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Old 10-08-2019, 08:53 AM
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Johanovich Johanovich is offline
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Default Re: Mantella baroni introgression? Very picture heavy!

Quote:
Originally Posted by hypostatic View Post
haha, yeah, there's still lots of old school genetics that gets done

The only real drawback for this approach is the fidelity of the available information. The paper is behind a paywall for me, so I'm not sure how rigorous their data collection methodology is. That said, the BLAST algorithm should prevent non species-specific primer binding. The primer design past is based on Primer3, which is also pretty accurate and trusted.

In this case a real positive control would be hard to get ahold of, due to the lack of information. But a well-designed assay would go a long way. You could start with just one individual to test things out, and run the baroni/madagascariensis primer sets in different tubes to avoid potential complications. And then as long as you only really get an amplicon of the expected size, you should be good.

OH and I guess the other thing you'd need is some sort of kit for amplifying genomic DNA from swabs or other similar method.

But yeah, this would be a nice thing for us to have in the hobby, to confirm/prove the identities of baroni/madagascariensis. Other methods are completely subjective...
Already had some sterile swabs available, so I swabbed their skin yesterday and extracted DNA from it (we had a few spare DNA extraction kits from free testers). I'll try and measure DNA content of the extracts and do a PCR test with general eukaryotic primers to see if the extractions worked. Hopefully most will work because I'd prefer to not have to swab them again, it's not exactly pleasant for them.

I'll try to get a hold of one of the primer pairs for each species. I've selected pairs which have different amplicon size for each species (618 bp for mad and 1028 bp for baroni), GC clamps and relatively low self-compatibility so hopefully they work. Won't able to get and test them before the end of the week, but I'll keep you posted when I get any results.
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