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Biology Articles » Methods & Techniques » Fast and non-invasive PCR sexing of primates: apes, Old World monkeys, New World monkeys and Strepsirrhines » Discussion

- Fast and non-invasive PCR sexing of primates: apes, Old World monkeys, New World monkeys and Strepsirrhines

In this study we have established a simple, accurate and widely applicable method for determining the sex of primate DNA samples by using triple primer PCR of a small region of the UTX/UTY gene. The ubiquitously transcribed tetratricopeptide repeat protein gene (UTX/UTY) is located on the X and Y chromosomes and our analysis identified a region in the human UTY having the highest identity to the mouse Y chromosome. Due to the high conservation of this region the triple primer PCR setup works in all primate species tested. Furthermore, the method contains an internal positive control (the shared primer), but should always be tested in samples of known sex before actual analysis is carried out. Also, it may be necessary to perform species specific optimization of annealing temperature and/or primer concentrations prior to analysis.

Since the nature of the Y chromosome allows deletion of many regions, it can be expected that with an increased number of individuals tested deletion of UTY might be found in low frequency (as reported for amelogenin-Y [15]). Furthermore, primer region mutations may result in non-identification of males due to PCR failure, which are known in the amelogenin system [9-11]. Generally, non-identification of males may result from non-amplification of the Y fragment and it has been argued [13] that all "females" (XX or XY with no Y amplification) should be analysed with at least two independent sex tests. It is now possible to combine the analysis of amelogenin, SRY and UTX/UTY in parallel for most primate species, thereby obtaining reliable sex determination (e.g. [7] and this study) using independent PCR runs. Given the low amount of DNA usually obtained from non-invasive sampling, the optimal approach for molecular sexing would be to run a single multiplex PCR analysis, but such a protocol has not been developed yet.

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