# Morph Market to offer DNA testing



## Malc (Oct 27, 2009)

RGI Genetic Tests for Reptiles - MorphMarket







www.morphmarket.com





A small piece of shed is all RGI needs to know if your animal is male or female, or if it contains certain morphs.

Wouldn’t you want to:


Save years of time by immediately proving your breeders?
Increase your sales by selling Pos Hets as 100% Het?
Verify that your purchases were advertised correctly?
Discover genes in your lines that you didn’t even know about?
Distinguish between incomplete dominants like Asphalt/Gravel/YB?
Find out if your special pet is a boy or girl?
Tests already available include:


Ball Python Morph Identification for Clown, Piebald, and Lavender Albino
Ball Python Sex Determination
Colubrid Sex Determination
Venomous Sex Determination
No idea of pricing yet, or if the test will identify all genes or just the recessive ones.... but if it's affordable it could take out a lot of the guesswork and disappointment for those working with certain morphs


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## Malum Argenteum (5 mo ago)

https://raregeneticsinc.com/


is down right now, but I recall they had prices listed.

Though I couldn't find any clear regulation on this, it seems that CITES appendix II requires permits for skins, skin pieces and skin scraps based on export data for those items for _P. regius_. Challenging hurdle for those keepers not in the US.


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## ian14 (Jan 2, 2008)

Malum Argenteum said:


> https://raregeneticsinc.com/
> 
> 
> is down right now, but I recall they had prices listed.
> ...


You are correct. Skins are a derivative. All pythons and boas are at the very least Appendix II. Whether a shed skin would be deemed a derivative or a waste product I am not sure.


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## Central Scotland Reptiles (Nov 23, 2008)

ian14 said:


> You are correct. Skins are a derivative. All pythons and boas are at the very least Appendix II. Whether a shed skin would be deemed a derivative or a waste product I am not sure.


You beat me to mentioning this potential issue.


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## Shellsfeathers&fur (Jan 18, 2009)

Can you sue them if they get it wrong?!


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## ian14 (Jan 2, 2008)

Shellsfeathers&fur said:


> Can you sue them if they get it wrong?!


Good point. 
My concern is how reliable this is. Unless a reputable genetics lab has mapped the genome of every known royal python morph then how can they guarantee an accurate result? We all know the mess that corn snake genetics are, and with how highly inbred royals now are, ots highly likely that many will be like corns with unknown and unexpected hidden genes.
So I'm not entirely sure how possible this is.
Legally too, if the shed skins are deemed a derivative, you can only send them via specified UK ports, you cannot just bung them in the post. You will need CITES export permits, and have the items checked at a designated port by Border Force to confirm that the specimen/s are as declared on the export permit.
It's not clear when these tests will actually be available.
When I'm back at work from sick leave I'll ask Border Force if they would consider a shed skin to be a derivative.


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## Elly66 (Feb 27, 2021)

Well, the website is very lacking in actual scientific information and I can't even get it to show "tests available ". Genetics is such a complex area and I'd certainly want to see a lot more proven detail before even considering it. That's before even looking at the legal restrictions of sending the shed.


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## Malc (Oct 27, 2009)

Some interesting points raised here. I don't fully understand how the science works, but presumably each gene will have its own "address", so you would look to detect if a gene is present at set addresses on the DNA strand, rather than set up a database of genes as this would need a database of base single gene examples.

Also interesting points regarding animals on CITIES, especially if this service is operated from outside the UK. I like the idea, but don't get the feeling it's been implemented well. Also as  mentioned, how would they stand if they misreported and found themselves in court


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## Malum Argenteum (5 mo ago)

The "science" looks pretty straightforward.









A community-science approach identifies genetic variants associated with three color morphs in ball pythons (Python regius)


Color morphs in ball pythons ( Python regius ) provide a unique and largely untapped resource for understanding the genetics of coloration in reptiles. Here we use a community-science approach to investigate the genetics of three color morphs affecting production of the pigment melanin. These...




www.biorxiv.org













A nonsense mutation in TFEC is the likely cause of the recessive piebald phenotype in ball pythons (Python regius)


Captive-bred ball pythons ( Python regius ) represent a powerful model system for studying the genetic basis of colour variation and Mendelian phenotypes in vertebrates. Although hundreds of Mendelian phenotypes (colour morphs) affecting colouration and patterning have been described for ball...




www.biorxiv.org





Sequence a bunch of animals that are known to have the gene, and then see what variants in the code stick out; do some predictive testing to validate the test; run some statistical analyses to show the results aren't due to chance. 

I won't pretend to know the actual technological details, but this sort of thing is the basis of PCV testing (which in the US, and I think the UK, is available at the hobbyist level for a wide range of pathogen testing for very low costs) and all the phylogenetic work that is being done in taxonomy lately (sequencing animals to see how closely they're related evolutionarily by looking at the difference between known genetic markers).

Not sure how the legal liability would be different than any other similar (or even technologically different) service, not whether there would be any hypothetical case that would be worth pursuing.


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## ian14 (Jan 2, 2008)

I think the concern is that what is being marketed is a genetic test on a know species to identify a specific mutant gene. That's different to identifying a pathogen, as that simply involves identifying the species.
Given the number of known mutant genes in royals, the only way, the absolute only way, they can achieve this is by taking confirmed morphs and sequencing the entire DNA to locate the locus at which the mutant gene sits AND identify the exact DNA sequence for the mutant gene causing that morph. 
They will then have to take the DNA of the specimen being examined, extract the sequence and compare to all the known mutant genes in order to identify what hidden genes it is carrying.
This is why I am rather dubious
DNA sexing isn't new, it's been used for parrots for a very long time, so that's not a concern. But identifying morph genes? Not so sure.


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## Malum Argenteum (5 mo ago)

ian14 said:


> Given the number of known mutant genes in royals, the only way, the absolute only way, they can achieve this is by taking confirmed morphs and sequencing the entire DNA to locate the locus at which the mutant gene sits AND identify the exact DNA sequence for the mutant gene causing that morph.
> They will then have to take the DNA of the specimen being examined, extract the sequence and compare to all the known mutant genes in order to identify what hidden genes it is carrying.
> This is why I am rather dubious


The procedure you describe in the first sentence is exactly what was done in the studies I linked.

"We used whole-genome sequencing of pooled samples followed by population genetic methods to delineate the genomic region containing the causal gene."

The procedure you describe in the second sentence is basically what PCR testing (for phylogenetic studies and pathogen testing) does. It doesn't sequence the whole genome of the sample, though, but rather amplifies certain regions of interest and then compares those to published genetic data (that amplification is the source of much of the false negatives in PCV testing, since the scale of the amplification can cause imperfect replication). The whole genome doesn't need to be compared, since the mutant locus is known. 

Except for knowing which sequences at which locations are the bases of the various morphs, none of this is novel as far as I can see.


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## ian14 (Jan 2, 2008)

Malum Argenteum said:


> Except for knowing which sequences at which locations are the bases of the various morphs, none of this is novel as far as I can see.


And that's my point!
Unless they have taken DNA from every known morph, sequenced it and identified the locus of each gene causing the mutation for the morph, and identified the DNA sequence of that mutant gene, then how on earth can they provide a DNA testing service?


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## Malum Argenteum (5 mo ago)

They're (planning to) provide the service for those morphs for which there are published sequences, and only for those morphs. This is stated in the MM link.

There's no reason that the genetic markers for every known morph must be published, any more than PCR testing for e.g. IBD Arenavirus requires that every known virus be sequenced. The testing will be 'positive/negative' testing for available morphs (currently Clown, Piebald, and Lavender Albino), not a printout of all the morphs a snake carries. 

Again, this is just like pathogen testing -- a sample is tested for whatever pathogen the client specifies, and if the test finds DNA for that pathogen, the test is positive, if not, negative. A PCR pathogen test doesn't produce a list of all the pathogens in the sample (though testing for each known pathogen could be done, of course; this would be one test per known pathogen).


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## StuG (Nov 4, 2009)

ian14 said:


> And that's my point!
> Unless they have taken DNA from every known morph, sequenced it and identified the locus of each gene causing the mutation for the morph, and identified the DNA sequence of that mutant gene, then how on earth can they provide a DNA testing service?


They only offer the service on morphs that they have done that with-Piebald, Clown and lav albino so far I think. There’s a couple if U.K. breeders that have sent skins over for poss het Lav clutches


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## ian14 (Jan 2, 2008)

StuG said:


> They only offer the service on morphs that they have done that with-Piebald, Clown and lav albino so far I think. There’s a couple if U.K. breeders that have sent skins over for poss het Lav clutches


That makes sense. It did seem very, very over-optimistic for this to be done for every morph! 
It does seem a good idea, I guess over time they will expand the morphs they can test for.
There are, I fear, still legal issues with sending shed skins for testing as they are being used for a commercial purpose. As I said, I will try to get an answer on this.


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## Malc (Oct 27, 2009)

ian14 said:


> That makes sense. It did seem very, very over-optimistic for this to be done for every morph!


from their website



> But this is just the start! 3-5 additional Ball Python morph tests will be added in the next few weeks. We are currently working on Ball Python Morph Identification for Desert Ghost, Hypo, Super OD/OD, Ultramel.
> More morph tests will be available each month.





ian14 said:


> And that's my point!
> Unless they have taken DNA from every known morph, sequenced it and identified the locus of each gene causing the mutation for the morph, and identified the DNA sequence of that mutant gene, then how on earth can they provide a DNA testing service?


So if I understand that correctly, its' not as if the location for the gene is in a set location in the DNA for every morph, the location could be anywhere in the full DNA code for that snake hence the need to sequence the full DNA.

I'm guessing the sex is easy as you only have two options and presumably they are at a known point in the sequence.

I'm also guessing that if a breeder has a test done to confirm that snake is a Pastave, enchi, super OD het for desert ghost or whatever, that will only push up the pricing as the breeder will want to cover the cost and provide the certification for that snakes "pedigree" (in effect). 

I seem to hear echoes of Theranos here ---- Ms Holmes hasn't escaped and jumped bail to set up a new company has she


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## ian14 (Jan 2, 2008)

Malc said:


> from their website
> 
> 
> 
> ...


That's it exactly.
A mutated gene that prevents melanin being produced will be in a different place in the DNA sequence to a mutated gene that causes, for example the spider effect, which will again be different to where the mutated gene sits that causes blue eyes, or black eyes, or pastel etc etc.
And yes, you can guarantee that the cost of that test will be passed on, thus upping the price of a het animal.
Amazing this has now been developed as the price of royals is dropping, or is that me being cynical?


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## Malum Argenteum (5 mo ago)

Well, PCR DNA tests for herp pathogens are $20 (RNA viruses are $25). Once morph testing gets established, there is no reason it would cost any more. 

A breeder who has produced a possible het and tests it to "prove out the het" stands to gain a lot more than $20 in most cases -- they can charge het price where without the test they could not. The availability of testing will also drive down the market price of 'possible hets' since there won't be a reason to buy a possible het for those morphs that are testable. Seems a financial benefit for the buyer here, not the seller.


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## Elly66 (Feb 27, 2021)

Testing for one specific, known gene is fairly simple.
DNA testing for the sex is probably the easiest test.

What makes genetic testing complex is that each chromosome has many layers - macro and micro chromosomes too start with. Each microchromome is likely to have it's own karyotype. Just one change from the norm in the karyotype can have a massive impact. 

Firstly, scientists need to map a huge amount of DNA sequences from the various species to get the norm. They need this before they can look at variations. Getting that norm isn't easy where many changes are being seen. I'd imagine testing wild snakes is the best way to find it, nature doesn't interfer as much as humans do. 

I'd certainly want to see scientific proof of this before spending money on a test and know just how specific the testing is. Personally, I'd be more interested in the health aspect of the snake and how much human intervention has affected them. If our want of morphs is causing detrimental issues to the snakes, then this science could be a wake up call.


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## StuG (Nov 4, 2009)

I think the cost of the test will be inconsequential compared to the value added by the presence or absence some of the hets.

It will help breeders with sales and buyers with certainty that the hets they are paying for are present.

I don’t know if they offer the service for other species but it would great to be able to but say a locality corn such as an Okeetee and be certain that various hets weren’t going to thrown out


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## Malum Argenteum (5 mo ago)

Elly66 said:


> I'd certainly want to see scientific proof of this before spending money on a test and know just how specific the testing is.


You might read the articles I linked. They are very descriptive. For example, here's the specificity of Lav Albino:

"Further test amplifications revealed that the Lavender Albino animal was homozygous for a 1,514-bp deletion spanning coding region 18 (Figs 3A and 3B). This deletion removes 36 amino acids from the protein and likely introduces a frameshift into the transcript, given that the coding regions downstream of the deletion are out of frame compared to coding regions upstream of the deletion."

There's a lot of explanation of how they found the code for the mutation. It isn't nearly so uncertain and difficult as you make it out to be; this sort of work is being done all over biology right now -- most of what I read on this is in taxonomy (mutations are used to date most recent ancestors). Here's a cool recent one on captive leo lines, for example. 

These aren't 1980's techniques, so the "karyotyping" considerations simply aren't relevant. Your assumptions about mapping aren't true, either; evolutionary conservation of genes makes it possible to look for a mutation in the same region that mutation has been found in other species' genomes. Wingless (as in fruit flies) is conserved in humans, for example, and all the data and findings about it is published and catalogued for other researchers to use.


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## Elly66 (Feb 27, 2021)

Malum Argenteum said:


> You might read the articles I linked. They are very descriptive. For example, here's the specificity of Lav Albino:
> 
> "Further test amplifications revealed that the Lavender Albino animal was homozygous for a 1,514-bp deletion spanning coding region 18 (Figs 3A and 3B). This deletion removes 36 amino acids from the protein and likely introduces a frameshift into the transcript, given that the coding regions downstream of the deletion are out of frame compared to coding regions upstream of the deletion."
> 
> ...


I will look at the links, along with my own research.

However, I disagree with your view on karyotyping being irrelevant and my " assumptions " on mapping not being true. Genetics is something I've had to look at up close and personally, so much so that I decided to study it at university in recent years. My eldest studied microbiology, so we have some interesting conversations. 

Mutations can develop anywhere and unique or previously unknown ones are only found by mapping the whole sequence and having the norm to compare it with. Yes, looking for a known mutation in certain areas of the sequence is relatively simple though. 

As I stated, my interest in snake genetics would me about how the mutations affect their wellbeing, especially when 2 snakes with different mutations are bred together. "Is the human want of morphs causing health problems?" is definitely more important to me.


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## ian14 (Jan 2, 2008)

Well this is now up and running, albeit for a fairly restricted list of morphs. They do state that owners from outside the US can use the service IF they can send skins legally.
I have contacted APHA for clarity as to whether a shed skin is a derivative part as the CITES Appendices and site do not make this clear.
Personally, I would strongly advise that those outside the US do NOT make use of this service until this issue is resolved as the penalties can be quite significant if you find yourself the wrong side of the law.
As soon as I get a reply I will post it here.


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## ian14 (Jan 2, 2008)

Well I've had a reply and I'm afraid it's bad news:

*Thank you for your email .



I have consulted with management and we would consider that snake skins require permits if they are CITES-listed, because the remit for CITES is that ‘all parts and derivatives’ are included in the rules. So, if the species appears on www.speciesplus.net as Annexe A, B or C then permits are required. The designated ports of exit and extry in/out of the UK are here: :

https://www.gov.uk/guidance/trading-cites-listed-species-through-uk-ports-and-airports-after-brexit



I hope this answers your question sufficiently, if not, please get back in touch.*

So in short, if you want to use this service you will need CITES permits and the skins can only be sent via a designated exit port.


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## Malc (Oct 27, 2009)

ian14 said:


> Well I've had a reply and I'm afraid it's bad news:
> 
> *Thank you for your email .
> 
> ...


Ian, did you make it clear that we're talking about a sloughed snake skin and not a skinned animal. It makes no sense as there is no trade or use for a normal shed


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## ian14 (Jan 2, 2008)

Malc said:


> Ian, did you make it clear that we're talking about a sloughed snake skin and not a skinned animal. It makes no sense as there is no trade or use for a normal shed


I did, yes. I have also asked for them to confirm that they are satisfied that the skin shed by a snake as it grows is controlled as a part or derivative under CITES.
The issue is that there is no requirement for there to be any value, the wording is "any part or derivative".
As it stands the current position from APHA, as the management authority for animals in the UK for CITES is that a shed skin is still a part or derivative and so is controlled.
As these tests require sending a skin to the US, you are therefore exporting a CITES specimen and so permits and other legal requirements kick in.


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## Malc (Oct 27, 2009)

ian14 said:


> I did, yes. I have also asked for them to confirm that they are satisfied that the skin shed by a snake as it grows is controlled as a part or derivative under CITES.
> The issue is that there is no requirement for there to be any value, the wording is "any part or derivative".
> As it stands the current position from APHA, as the management authority for animals in the UK for CITES is that a shed skin is still a part or derivative and so is controlled.
> As these tests require sending a skin to the US, you are therefore exporting a CITES specimen and so permits and other legal requirements kick in.


Thanks for the clarification. Not expecting it to be high numbers, but I bet someone will get caught out by this.


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