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Investigation of RAPDs and microsatellites for use in South African cranes.King, Heather Anne. 29 November 2013 (has links)
The three South African crane species, namely, the Wattled Crane (Bugeranus
carunculatus), the Blue Crane (Anthropoides paradisea) and the Grey Crowned Crane
(Balearica regulorum regulorum) are all threatened. South African legislation protects the
cranes, however eggs and/or fledglings are sometimes illegally collected from the wild. These
are then sold, often by registered breeders, who falsely claim them as the offspring of their
captive breeding pair. DNA fingerprinting is one method to detect this crime.
Fifteen RAPD primers were screened for polymorphism in the three species. Seven
primers produced polymorphic profiles in the Blue Crane and eight each in the Grey Crowned
Crane and Wattled Crane, with an average of 14.57, 12.38 and 5.88 scorable loci per primer,
respectively. The Band Sharing Coefficient for unrelated individuals was found to be 0.665,
0.745 and 0.736 for the Blue, Grey Crowned and Wattled Crane respectively.
Five microsatellite primers, originally developed for use in Whooping Cranes (Grus
american), had previously been shown to be polymorphic in the Wattled Crane. This was also
the case in this study with an average of 3.6 alleles per primer. Although all primers cross
amplified, only a single primer each showed polymorphism in the Blue Crane (showing 6
alleles) and the Grey Crowned Crane (showing 5 alleles).
The RAPDs were found to be irreproducible, show high numbers of novel bands and
had parent: offspring BSC values that were not significantly higher than those of unrelated
individuals. Statistics showed that, in the Blue Crane, the probability that misassigned parents
would be detected was low whilst there was an almost certainty that true parents would be
incorrectly excluded.
The five microsatellite primers examined gave exclusionary powers of 0.869 and 0.641
where one or two parents were unknown in the Wattled Crane. The exclusionary powers for
the Blue Crane and Grey Crowned Crane calculated at only one locus were much lower. It was concluded that RAPDs were totally inappropriate for parentage analyses,
however, microsatellites are a suitable technique and recommendations are made that other microsatellites, developed for other species of crane, should be examined for their potential in
this respect. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2004.
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