Summary
Gender identification of forensic samples was determined by amplifying a segment of the X-Y homologous gene amelogenin. Using a single pair of primers spanning part of the first intron, 106 by and 112 by PCR products were generated from the X and Y homologues respectively, which were then resolved by agarose gel electrophoresis. This test enabled as little as 20 pg of DNA from severely degraded bones to be amplified and typed in a single tube reaction. Furthermore, using dye-labelled primers, it was possible to quantitate, by automated fluorescence detection, the relative yields of X and Y -specific PCR products generated from mixtures of male and female DNA. The versatility of this sex test was further demonstrated by co-amplifying with the HLA-DQA1 Amplitype kit in a combined gender/identity DNA test.
Zusammenfassung
Durch Amplifikation eines Segments des X-Y-homologen Gens Amelogenin wurde eine Geschlechtsidentifikation forensischer Proben durchgeführt. Mit einem einzigen Primerpaar, welches einen Teil des ersten Introns überspannt, wurden PCR-Produkte mit 106bp und 112bp von den homologen Anteilen des X- und des Y-Chromosoms generiert, welche dann mit Hilfe der Agarosegelelektrophorese aufgetrennt wurden. Dieser Test erlaubte es, daß so geringe Mengen wie 20pg DNA von stärkergradig degradierten Knochen amplifiziert und ineiner Einzelreaktion typisiert wurden. Durch Benutzung von farbstoffmarkierten Primern war es ferner möglich, durch automatisierte Fluoreszenzdetektion die relative Ausbeute von X- und Yspezifischen PCR-Produkten zu quantifizieren, wie sie von Mischungen männlicher und weiblicher DNA generiert wurden. Die Vielseitigkeit dieses Sex-Tests wurde ferner dadurch nachgewiesen, daß eine Co-Amplifikation mit dem HLA-DQA1 „Amplitype”-Kit in einem kombinierten Geschlechtsbestimmungs-und Identifizierungs-DNA-Test möglich war.
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Mannucci, A., Sullivan, K.M., Ivanov, P.L. et al. Forensic application of a rapid and quantitative DNA sex test by amplification of the X-Y homologous gene amelogenin. Int J Leg Med 106, 190–193 (1994). https://doi.org/10.1007/BF01371335
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DOI: https://doi.org/10.1007/BF01371335