Multiplex primer extension assay for identiﬁcation of paternal lineages in domestic goat (Capra hircus)
The IDENTIFICA SNPYGoat Kit is a rapid and robust multiplex PCR/primer extension assay to genotype a set of goat Y chromosome single nucleotide polymorphisms (SNPs). Four diagnostic SNPs located in two ﬂanking domains of the SRY gene (promoter region and 3 untranslated region) were selected to allow the discrimination of the four Y-chromosomal haplotypes Y1A, Y1B, Y1C and Y2 identiﬁed so far in domestic goat populations.
The detection of SNP variants is based on the SNaPshot single-base sequencing (also known as minisequencing), which consists in using dideoxynucleotides (ddNTPs) for singlebase extension of an unlabeled primer that anneals one base upstream to the relevant SNP.
An auxiliary software was developed to simplify the process of haplotype identiﬁcation directly from the sample proﬁle.
|Primer set A (10x)
Positive Control 1 (Capra hircus control DNA)
|25 reactions (K007AP)
50 reactions (K007BP)
|SNaPshot single-base sequencing
Pereira F, Carneiro J, Soares P, Maciel S, Nejmeddine F, Lenstra JA, Gusmão L, Amorim A.
A multiplex primer extension assay for the rapid identification of paternal lineages in domestic goat (Capra hircus): laying the foundations for a detailed caprine Y chromosome phylogeny.
Molecular Phylogenetics and Evolution. 2008. 49:663-668.
Pereira F, Queirós S, Gusmão L, Nijman IJ, Cuppen E, Lenstra JA, the Econogene Consortium, Davis SJM, Nejmeddine F, Amorim A.
Tracing the history of goat pastoralism: new clues from mitochondrial and Y chromosome DNA in North Africa.
Molecular Biology and Evolution. 2009. 26:2765-73.