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De Gruyter, Clinical Chemistry and Laboratory Medicine, 2(50)

DOI: 10.1515/cclm.2011.758

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Alternative methods to a TaqMan assay to detect a tri-allelic single nucleotide polymorphism rs757210 in the HNF1β gene

This paper is available in a repository.
This paper is available in a repository.

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Abstract

AbstractSeveral studies report difficulties in genotypingPyrosequencing and high resolution melting analysis of small amplicons (HRM) were developed and tested in panels of type 2 diabetes mellitus patients (n=258) and healthy blood donors (n=183). Results were confirmed by Sanger sequencing.With pyrosequencing, allele frequencies for the A, G and C allele of 0.42, 0.56, 0.02 and 0.37, 0.62, 0.01 were established in the panel of type 2 diabetes mellitus patients and healthy blood donors, respectively. Similar results were found using the more routinely available HRM method. Results for pyrosequencing and HRM were in 99.6% concordance.Pyrosequencing and HRM can be used to genotype the tri-allelic SNP rs757210 in the