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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.
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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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to main sexing page
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