Abstract
BackgroundSimple and precise methods for sex determination in animals are a pre-requisite for a number of applications in animal production and forensics. However, some of the existing methods depend only on the detection of Y-chromosome specific sequences. Therefore, the abscence of a signal does not necessarily mean that the sample is of female origin, because experimental errors can also lead to negative results. Thus, the detection of Y- and X-chromosome specific sequences is advantageous.ResultsA novel method for sex identification in mammals (sheep, Ovis aries and European red deer, Cervus elaphus) is described, using a polymerase chain reaction (PCR) and sequencing of a part of the amelogenin gene. A partial sequence of the amelogenin gene of sheep and red deer was obtained, which exists on both X and Y chromosomes with a deletion region on the Y chromosome. With a specific pair of primers a DNA fragment of different length between the male and female mammal was amplified.ConclusionPCR amplification using the amelogenin gene primers is useful in sex identification of samples from sheep and red deer and can be applied to DNA analysis of micro samples with small amounts of DNA such as hair roots as well as bones or embryo biopsies.
Highlights
Simple and precise methods for sex determination in animals are a pre-requisite for a number of applications in animal production and forensics
When the results are negative from amplification of only the SRY gene, it cannot be assumed that the individual is female or that there was a mistake in the experimental process
Besides a few minor single nucleotide ins/dels, the most prominent ins/del is between positions 63 and 107 (45 bp) and positions 127 and 136 (10 bp) according to Fig. 1. This major ins/del leads to a shorter amplicon that can be detected in an agarose gel
Summary
Simple and precise methods for sex determination in animals are a pre-requisite for a number of applications in animal production and forensics. Some of the existing methods depend only on the detection of Y-chromosome specific sequences. The detection of Y- and X-chromosome specific sequences is advantageous. The male is identified by amplifying the SRY gene (sex-determining region Y) which is a Y chromosome-specific sequence. When the results are negative from amplification of only the SRY gene, it cannot be assumed that the individual is female or that there was a mistake in the experimental process. In sex identification of ursides by PCR, primers that amplify the SRY gene together with the mitochondrial DNA control region or the ZFX/Y region have been used [1,2]. Recent reports have described the use of low-stringency PCR or the detection of X/Y specific restriction fragment polymorphisms (RFLP) in sheep and goats [3,4]. The amelogenin (AMEL) gene, which exists on both X and Y chromosomes, has been used to determine (page number not for citation purposes)
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