Advances in Forensic Haemogenetics pp 329-332
DNA Sex Test: A New Rapid and Quantitative Forensic Approach Using Amelogenin Gene Based Fluorescent PCR
- Cite this paper as:
- Mannucci A., Sullivan K.M., Ivanov P.L., Kimpton C., Gill P. (1994) DNA Sex Test: A New Rapid and Quantitative Forensic Approach Using Amelogenin Gene Based Fluorescent PCR. In: Bär W., Fiori A., Rossi U. (eds) Advances in Forensic Haemogenetics. Advances in Forensic Haemogenetics, vol 5. Springer, Berlin, Heidelberg
For forensic PCR applications, it is recommended that the investigation of X and Y sequences should be carried out in parallel or simultaneously, and the distinction of male and female DNA cannot be made based solely on the absence of a band . Several PCR-based tests have been developed for gender identification, among them multicopy repeat sequences on the X and Y chromosomes can be amplified together and provide a highly sensitive assay but quantitation of the relative X/Y product is not possible because of significant differences in repeat copy number [2–4]. Alternatively, single copy X-Y homologous regions such as amelogenin offer the advantage of requiring only one pair of primers and both X and Y sequences are of equal copy number . Primers described by Sullivan and coll. flank a 6bp deletion within intron 1 of the X homologue resulting in 106bp and 112bp PCF, products from the X and Y chromosomes respectively . Dye labelled PCR products were generated using one primer coupled with fluorescent dye ‘FAM’ via a 5’ aminolinker. These primers allowed DNA samples ranging from 10pg to 100ng to be amplified through 35 cycles comprising 1 minute at 94°C, 1 minute at 60°C and 1 minute at 72°C in a Perkin Elmer 480 thermal cycler. PCR products (20 μl) were visualised simply after electrophoresis in a 4% agarose gel for 90 mins at 100V and by ethidium bromide staining.
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