Quantification of specific mutation with HRM for regions of interest

Robert Loewe
GeneWake GmbH, Germany

HRM has been implemented on numerous instruments with many chemistry options. Despite this fact optimization and quantification of e.g. mutational status has not yet reached most of the labs. Therefor a comparison of instruments and different chemistry was done. As a model system the SNP responsable for lactose intolerance (rs4988235) was used to find optimal amplicon length (tested from 60 – 653bp). Also we tested the possibility to calculate the mutational percentage for diagnostic purposes. The outcome presented a potential solution for quantitative HRM with the possible usage in screening minute sample material for mutational status.

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