DNA tests for the blue egg gene, shipping samples to Germany

[DarJones]

I will be shipping a package of feathers to Justus Liebig University in Germany Monday for DNA tests for the blue egg gene (oocyanin). Here are the steps and costs so far. These steps are a bit convoluted and I am going to post in detail so be prepared for a LONG set of steps. I am taking pictures and will post them in a few days when I have time to get them uploaded to my website.

Samples have to be packed in foil pouches and heat treated to 320F for 30 minutes. Then they have to be packed and shipped to Justus Liebig University to the veterinary department professor. A lab assistant will then unpack the samples and begin tests. Results will be returned via email.

Costs:
1. Time to gather the feather samples is about 3 minutes per bird sampled
2. $.05 cost for foil each to pack the feather samples
3. $.05 each for ziploc bags for temporary storage of the feather samples
4. $1.25 X 2 for heat sensitive labels to prove the samples are heat sterilized.
5. $.25 estimated for propane to heat my stove to 320F for 30 minutes
6. $? for shipping, estimated to be about $30 for an international small package
7. $31.72 (€28) quoted cost for each sample processed

Taking feather samples was fairly easy. I waited until the chickens were on the roost, then went in with vinyl gloves, a box of ziploc bags, a sharpie marker, and legbands. Wearing gloves, I caught a chicken by the legs and then held it head down with feet pointing away from me so I could easily pull out about 6 breast feathers. Ideal size feathers are about 4 inches long. Apply a numbered legband, place the feather sample in a ziploc bag, return the chicken to the roost, then label the ziploc bag with the legband number and descriptive information. I could do the entire process in about a minute once I had everything in place. The gloves are to avoid DNA contamination.

I carried the ziploc bags indoors and set up a work area to pack the feather samples in foil pouches. I put on a fresh pair of vinyl gloves (did not want any DNA contamination from the gloves I had been using) and started preparing foil sheets. I tore off a piece of aluminum foil (Reynolds heavy aluminum foil) about 6 inches long by 14 inches wide and then cut it in half making two pieces each about 6 inches by 7 inches. I then folded the foil about an inch and a half along one edge to make a V for the feathers. I pulled a ziploc bag and moved feathers into the foil V and then folded the foil 2 more times making a flat strip about 1.5 inches wide by 7 inches long. Then I folded the ends over about 3/8 of an inch completely enclosing the feathers. Using a wide point ink pen, I scored the legband number into the foil. Then I used a sharpie to mark over the number in black to make it highly visible. I was not sure whether the sharpie would survive heating in the oven, but I am certain the foil pouches will retain the number pressed into the foil.

I then pulled off a piece of aluminum foil about 14" by 14" and placed the group of individual pouches on it and made a large pouch around the group of smaller pouches. When finished it was about 8 inches long by 4 inches wide and 1/2 inch thick. I could have placed about 20 small pouches in each large pouch. In this case, I put 12 in one and 10 in the other. I applied 2 heat sensitive 320F labels to each pouch in such a way that the pouch could not be opened without removing the label. Then I heated my oven to 320F and placed the pouches in it for 30 minutes. At that time, all of the heat sensitive areas on the strip had changed to black. I removed the pouches and let them cool.

The rest was paperwork. I took the ziploc bags (organized by large pouch) and filled out an email to Justus Liebig University. The email contains the legband number plus descriptive information for each bird sampled.

JLU has been working on a test for the rose comb inversion on chromosome 7. They are going to test my samples to see how many are homozygous for rose comb. The test is not yet accurate enough for general use. I deliberately included a straight comb rooster along with 12 rose comb roosters in my samples. I can tell by looks that most of the roosters only have one copy of the rose comb inversion. If I am very very lucky, there will be a rooster that is homozygous for blue eggs and homozygous for rose comb.

Why use DNA tests that cost roughly $35 for each chicken sampled? Because it would take at least 4 times that much money plus at least a year of time to identify a homozygous rooster using test breeding. I am saving at least $1500 in feed and care costs plus a huge number of chicks to raise trying to find a rooster that is homozygous for blue egg.

 

[Amer]

That’s cool! Nice motive too. What’s your eventual goal for the project?

 

[SpicyDisaster]

Thank you for the write up! What breeds are your roos you are sending samples in for?

 

[DarJones]

My long term goal is to stabilize a blue egg laying Silver Laced Wyandotte. I am at the point where identifying a couple of homozygous roosters is critical to the breeding program. All of my birds currently look like Wyandottes though I still have about 4 golden laced birds. I'm going to remove two of them tonight because they have straight comb and one of them lays brown eggs. I need to cull about 6 chickens total (out of 47) that lay brown eggs. It takes time to identify the brown egg layers. I tagged two of them today.

 

[Sally PB]

Good luck with your project. Please update this thread as you get more information.

 

[3KillerBs]

 

[SpicyDisaster]

That sounds like an awesome project, can't wait to hear the results! What breeds did you use to bring in the blue egg gene? I've seen people mention cream legbars and Americauna so I'd be curious what you used and the rain behind your choice.

 

[DarJones]

I started with Silver Laced Wyandottes from Jerry Foley ( http://www.foleyswaterfowl.com/ ) and with blue egg laying Brown Leghorns from UARK back when Keith Bramwell was developing them. The reasoning is fairly simple. I needed a straight comb blue egg layer in order to combine blue egg with rose comb. Pea comb birds won't work because that produces walnut comb (Eliminates Ameraucanas). I thought hard about using Legbars but in a detailed study of the genetics found something just about everybody missed. Legbars carry a recessive gene that limits egg production potential (1 in 10 chance of a low egg producer) plus they mostly lay green eggs and I wanted blue eggs. I am friends with Glenn Drowns at Sandhill Preservation and asked him about possibilities for a cross. He had some Manx Rumpies that laid blue eggs and one bird was laced (huge advantage, already laced) but that would mean tying up the genetics with rumpless which I really did not want to do. I contacted Keith Bramwell and was told at first that he was not releasing birds at the time, but about 3 months later he emailed and offered me some eggs from his blue egg laying Brown Leghorns. Purely by chance, he was attending a conference in Atlanta and one of his grad students was going to drive on the interstate that passes less than 3 miles from my home. We met at the interstate and I picked up 3 flats (30 eggs each, 90 eggs total) and brought them home to put in my incubators. I had to agree not to sell or release the brown leghorns and only use them for crossbreeding as a condition of getting the eggs. There were other possibilities but overall the brown leghorns had the least genetic conflicts and gave the most advantages including high production of large sky blue eggs.

In the course of making crosses and studying the results, I learned a great deal about the genetics involved. For example, one bird showed up rumpless. Where did that come from? A few white egg layers showed up. This was expected as segregation could easily separate out the genes to turn off brown egg with no copies of the blue egg gene. I also had one hen that was yellow with speckles. Tell me how that comes from a brown leghorn crossed with silver laced Wyandottes. Crosses all laid very lightly tan tinted eggs. Why? It turned out that Brown Leghorns carry a gene known as "zinc white" which almost entirely suppresses porphyrin (brown eggs). Zinc white was a watershed moment for me as I realized I could leverage it in selecting for bright blue eggs. I can wipe an egg with a damp cloth and determine if it has the tan tint. If so, I know the hen that laid the egg has porphyrin turned on but blocked by zinc white. I just watch for hens that lay bright blue eggs because their offspring will tend toward pure blue eggs. Tons more I could post, but this is enough for now.

 

[SpicyDisaster]

I started with Silver Laced Wyandottes from Jerry Foley ( http://www.foleyswaterfowl.com/ ) and with blue egg laying Brown Leghorns from UARK back when Keith Bramwell was developing them. The reasoning is fairly simple. I needed a straight comb blue egg layer in order to combine blue egg with rose comb. Pea comb birds won't work because that produces walnut comb (Eliminates Ameraucanas). I thought hard about using Legbars but in a detailed study of the genetics found something just about everybody missed. Legbars carry a recessive gene that limits egg production potential (1 in 10 chance of a low egg producer) plus they mostly lay green eggs and I wanted blue eggs. I am friends with Glenn Drowns at Sandhill Preservation and asked him about possibilities for a cross. He had some Manx Rumpies that laid blue eggs and one bird was laced (huge advantage, already laced) but that would mean tying up the genetics with rumpless which I really did not want to do. I contacted Keith Bramwell and was told at first that he was not releasing birds at the time, but about 3 months later he emailed and offered me some eggs from his blue egg laying Brown Leghorns. Purely by chance, he was attending a conference in Atlanta and one of his grad students was going to drive on the interstate that passes less than 3 miles from my home. We met at the interstate and I picked up 3 flats (30 eggs each, 90 eggs total) and brought them home to put in my incubators. I had to agree not to sell or release the brown leghorns and only use them for crossbreeding as a condition of getting the eggs. There were other possibilities but overall the brown leghorns had the least genetic conflicts and gave the most advantages including high production of large sky blue eggs.

In the course of making crosses and studying the results, I learned a great deal about the genetics involved. For example, one bird showed up rumpless. Where did that come from? A few white egg layers showed up. This was expected as segregation could easily separate out the genes to turn off brown egg with no copies of the blue egg gene. I also had one hen that was yellow with speckles. Tell me how that comes from a brown leghorn crossed with silver laced Wyandottes. Crosses all laid very lightly tan tinted eggs. Why? It turned out that Brown Leghorns carry a gene known as "zinc white" which almost entirely suppresses porphyrin (brown eggs). Zinc white was a watershed moment for me as I realized I could leverage it in selecting for bright blue eggs. I can wipe an egg with a damp cloth and determine if it has the tan tint. If so, I know the hen that laid the egg has porphyrin turned on but blocked by zinc white. I just watch for hens that lay bright blue eggs because their offspring will tend toward pure blue eggs. Tons more I could post, but this is enough for now.

I'm going to have to come back and finish digesting this information a few times. Just really beginning my foray into genetics so I appreciate the wealth of information you've already provided. Sounds like you had a lot of serendipity on your side with the Brown Leghorns.

 

[Amer]

I started with Silver Laced Wyandottes from Jerry Foley ( http://www.foleyswaterfowl.com/ ) and with blue egg laying Brown Leghorns from UARK back when Keith Bramwell was developing them. The reasoning is fairly simple. I needed a straight comb blue egg layer in order to combine blue egg with rose comb. Pea comb birds won't work because that produces walnut comb (Eliminates Ameraucanas). I thought hard about using Legbars but in a detailed study of the genetics found something just about everybody missed. Legbars carry a recessive gene that limits egg production potential (1 in 10 chance of a low egg producer) plus they mostly lay green eggs and I wanted blue eggs. I am friends with Glenn Drowns at Sandhill Preservation and asked him about possibilities for a cross. He had some Manx Rumpies that laid blue eggs and one bird was laced (huge advantage, already laced) but that would mean tying up the genetics with rumpless which I really did not want to do. I contacted Keith Bramwell and was told at first that he was not releasing birds at the time, but about 3 months later he emailed and offered me some eggs from his blue egg laying Brown Leghorns. Purely by chance, he was attending a conference in Atlanta and one of his grad students was going to drive on the interstate that passes less than 3 miles from my home. We met at the interstate and I picked up 3 flats (30 eggs each, 90 eggs total) and brought them home to put in my incubators. I had to agree not to sell or release the brown leghorns and only use them for crossbreeding as a condition of getting the eggs. There were other possibilities but overall the brown leghorns had the least genetic conflicts and gave the most advantages including high production of large sky blue eggs.

In the course of making crosses and studying the results, I learned a great deal about the genetics involved. For example, one bird showed up rumpless. Where did that come from? A few white egg layers showed up. This was expected as segregation could easily separate out the genes to turn off brown egg with no copies of the blue egg gene. I also had one hen that was yellow with speckles. Tell me how that comes from a brown leghorn crossed with silver laced Wyandottes. Crosses all laid very lightly tan tinted eggs. Why? It turned out that Brown Leghorns carry a gene known as "zinc white" which almost entirely suppresses porphyrin (brown eggs). Zinc white was a watershed moment for me as I realized I could leverage it in selecting for bright blue eggs. I can wipe an egg with a damp cloth and determine if it has the tan tint. If so, I know the hen that laid the egg has porphyrin turned on but blocked by zinc white. I just watch for hens that lay bright blue eggs because their offspring will tend toward pure blue eggs. Tons more I could post, but this is enough for now.

I didn’t know about those genes for egg color and production. Interesting!