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 Dot-Blot DNA Sexing Procedure


Dot-blot assays for determining the sex of a variety of species of birds were developed in 1990 by Dr. Siwo de Kloet, a former genetics professor at Florida State University, and his wife Dineke de Kloet. Developed specifically to identify the sex of most species of parrots and parakeets, this method proved to be extremely reliable when sufficient amounts of high molecular weight genomic DNA could be collected.
Fortunately, unlike mammalian (or human blood) in which only the white blood cells contain a nucleus with DNA, avian blood contains nucleated red and white blood cells. Therefore, a single drop of avian blood contains much more DNA than a single drop of mammalian blood. This high concentration of DNA in avian blood allows us to offer a Dot-blot assay from as little as 20ul (single drop) of whole blood.

The dot-blot procedure used by Avian Biotech consists of the following steps: First, DNA concentrations of each sample is measured using a spectrometer to ensure the proper concentration of high molecular weight DNA. Next, a dot-blot apparatus is used to clearly place each DNA sample onto a nylon membrane. The membrane is then hybridized with a chemolumonesant, sex specific DNA probe. Finally, an X-ray film is exposed to the membrane illuminating the differences between male and female samples. Results are cross checked with DNA concentrations and controls to ensure the highest level of accuracy.





Fig.1 represents an actual scan of a Dot-blot result. The black dots represent female samples, and the blank circles represent male samples. A1, A2, A3, A4, and A5 are Umbrella Cockatoos; B1, B2, B3, B4, and B5 are Alexandrian Parakeets; C1, C2, C3, C4, C5, D1, and D2 are Congo African Greys; D3, D4, and D5 are Chattering Lories; E1, E2, E3,E4, and E5 are Quaker Parakeets; F1, F2, F3, and F4 are Blue & Gold Macaws; F5 and G1 are Senegal Parrots; and G2, G3, G4, and G5 are Yellow-naped Amazons. The dot-blot procedure is best suited for sexing most Psittacines when fresh whole-blood samples are submitted.

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Avian Biotech International
1336 Timberlane Road    Tallahassee, FL 32312-1766
850-386-1145 or 800-514-9672 (Office)  850-386-1146 (Fax)

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