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  Česky / Czech version Klin. Biochem. Metab., 13 (34), 2005, No. 2, p. 87–91.
 
Using LabMAP Luminex Technology for SNP Detection 
Bruchová H.1, Kráčmarová A.1, Černý V.2, Brdička R.1 

1Ústav hematologie a krevní transfuze, Praha 2Archeologický ústav AV ČR, Praha
 


Summary:

       One single nucleotide polymorphism (SNP) is estimated to occur approximately in every 1000 bp of the genome. So SNPs are the most prevalent genetic variations in human genome and they may serve as markers in genetic studies of human population. LabMAP Luminex technology represents a powerful array platform using fluorescent polystyrene microspheres as a solid support for oligonucleotide probes. The LabMAP Luminex system is capable to detect 100 different analytes simultaneously in a single sample. Company Marligen has developed SNP-Y Identification System for SNP profiling of the Y chromosome on Luminex instrument. The kit enables to determinate 43 SNPs including amelogenin marker to verify gender. Using the kit we tested 30 males from Southern Bohemia and Moravia (15 males from Jindřichův Hradec and 15 males from Třebíč). The selected males were born in the region as well as their fathers. Genomic DNA was isolated from peripheral blood of the males. Studied polymorphic loci were amplified by multiplexed PCR and labelled with fluorescent tag. Labelled PCR products were hybridized to allele-specific probes immobilized on beads. The bead mixes were analyzed by Luminex 100 System that measured integral intensity of sample fluorescence. The aims of the study were an optimalization of Luminex technology for SNP detection and a determination of haplogroups according to identified SNPs in males. The haplogroups may reflect origin and age of population, migration routes, history events etc. Out of 42 tested SNPs we identified different allelic frequencies in 10 polymorphisms. SNPs M45, M207 demonstrated similar frequencies of both alleles. In M42, M94, M168 we observed only derived allele. In the rest of the SNPs in all Y-chromosomes ancestral alleles were detected. Although in the Třebíč group we determinated 6 different haplogroups, in the Jindřichův Hradec group only 4 haplogroups were observed. However in both groups the predominant haplogroup was R that is typical for Europe and North-western Asia.

        Key words: Luminex, bead array, SNP, Y-chromosome, Třebíč, Jindřichův Hradec.
       

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