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DNA Paternity Testing

© By Cottonwoods Kennel
Previous printed version edited by Kim Miles

A breeder once told me that even though there were two males in a pen with one of his females, he could distinguish which was the sire just by looking at the pups. If faced with the possibility of two sires, what would you do? Could you tell which one was the sire? This year, something similar, yet unintentional, happened to us.
Mia   In the past, I would roll my eyes whenever I heard someone say they had an "accidental" litter.  I couldn’t help but think, “You have a female in heat... she's in with a male. Ok, where's the "accident?".
My thoughts have become a  little more "generous", or understanding because of what happened to us this year.
  Our animals normally breed during mid-February, so we separate the males and females during the last part of January. This has worked for many years with no mishaps. However, this year, on January 16th a commotion led me to the front kennel where Mia and Ash were tied. Mia had exhibited no signs of  bleeding, swelling or any indication of a willingness to mate.
We moved her immediately, calling our vet and asking him what our best options were. He advised against any medical action concerning the potential pregnancy. Although we anticipated that she might be pregnant, all we could do was simply hope that she was not — but prepare as if she was. We were pretty sure Ash, the alpha male, would be the sire sense we had seen him in a tie with Mia. From what we had observed, Maverick, the beta male, had shown no interest in Mia and he also seemed to be sufficiently cowed by the alpha. Although there was a chance that Maverick Ash could have bred Mia, all indications pointed to Ash.
 It was not long before we had our answer, Mia was definitely pregnant and we were going to be having our first “accidental” puppies. We were now faced with a big question: “Who’s the sire?” We just “knew” that it would be Ash, but couldn’t rule out Maverick, the second male in the pen, as a possible (or even a multiple) sire. Although we had not seen them mate, we couldn’t be 100% sure that such a mating did not occur; and while multiple-sired litters (i.e., litters with more than one father) are less common when it comes to these canines, they can and do happen. With our dogs we felt there was only one course of action: DNA profiling of the dam, possible sires and all of the puppies.

DNA PROFILING: The last five years has seen a sharp rise in DNA profiling for verifying paternity, and there are several labs to choose from, some of which are listed below. These labs send out swab test kits free of charge, along with detailed instructions. In the test kits, nylon bristle swabs for sample collection seem to be the most commonly used collecting tool, although foam swabs are also available.
  The sample collection method is non-invasive (i.e. no blood or hair bulb tissue is needed). The nylon bristle swab or the foam swab is inserted into the animal’s mouth between the gum and the cheek, where it collects loose check cells. The swab is then packaged and mailed back to the lab. DNA profiling is used on pets for identification and to verify ancestry. According to Genomic, a company specializing in DNA profiling, true parentage can be detected even within highly inbred lines to a reported accuracy of 99.8 %. Once the test is completed, a certificate is mailed to the owner, reporting the dog’s DNA profile in an alphanumeric code sequence and a color bar code.
  For Wolfdog owners: Our veterinarian assisted us in searching for a competent lab that would perform DNA profiling on Wolfdogs. He contacted one lab that immediately hung up on him after he mentioned the word “wolfdog.” However, I was a little more successful, the labs I called were very willing to accept them. 

List of Labs contacted concerning DNA testing


  Mia gave birth to five healthy puppies in March. One was black, like Maverick, and four were brown. Our puppies are born dark—and lighten in color as they age. Brown puppies generally become agouti or "grizzled" in color, which is the coloration of both Mia and Ash. My husband was sure that the black pup (#2) would be Maverick’s and that the remaining puppies in the litter would be Ash’s. On the other hand, I thought two pups would be Maverick’s (the black one and one other) and that the remaining puppies would be Ash’s. To our complete surprise, none of the puppies were sired by Ash -  even though we had seen Mia and Ash copulate. All of the puppies were sired by Maverick. (He must have been sneaking around in the dark :-) As a side note, one of the puppies  is the image of Ash!
  This only reaffirms my personal thoughts on the breeder’s comments posted at the beginning of this article; one can only question all of the pedigrees of puppies he produced with more than one male present during breeding season.
  Registering animals and documenting lineage is worthwhile, but this, alone, should not be considered “verified.” My situation has caused me to do a lot of thinking about taking advantage of the genetic testing services that are now available—both for identification, and for the assurance of my puppies new owners.


Two versions of genetic alleles are found at each site examined. One of the alleles is inherited from the sire, the other from the dam. Labs use letters of the alphabet to represent each known allele.

Parentage verification is done by checking to see if the letters at each site in the pup, can be formed by selecting one allele from each of the parents.

If a sire with alleles BG at the second marker mates with a dam with alleles PS at the second marker, they can produce offspring with alphabetic combinations at the second marker of:  BP, PG, BS, SG.

If the puppy in question has one of those combinations, it's called an inclusion
If the puppy does not have one of those combinations, it's called an exclusion


Excluded Male


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Cottonwoods DNA Pages



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