Introduction ore than 20 years have elapsed since the first single base pair substitution underlying an inherited disease in humans was characterized at the DNA level. Disease-associated gene lesions are currently collected and publicized by the Human Gene MUTATION Database (HGMD) in Cardiff, locusspecific MUTATION databases, and to some extent by the Genome Database (GDB) and Online Mendelian Inheritance in Man (OMIM) as comprehensive and up-to-date resources for information on genetic disorders and MUTATIONs causing them.1 – 4

Objective(s): Denaturing high performance liquid chromatography (DHPLC) is a high throughput approach for screening DNA sequence variations. To assess oven calibration, cartridge performance, buffer composition and stability, the WAVE Low and High Range MUTATION Standards are employed to ensure reproducibility and accuracy of the chromatographic analysis. The purpose of this study was to provide a cost-effective homemade MUTATION standard for DHPLC analysis.Materials and Methods: DHPLC was performed to evaluate different elution temperatures of a 374 bp DNA fragment with C>A MUTATION at position of 59 to achieve a peak profile similar to the Low MUTATION Standard. In order to verify the reproducibility of the homemade MUTATION standard using DHPLC, 15 different experiments were performed to compare the homemade MUTATION standard, the WAVE Low Range MUTATION Standard with a positive DNA control sample.Results: We identified a comparable elution temperature and a peak profile with the WAVE Low Range MUTATION Standard.Conclusion: This study confirmed the reproducibility of the peak profile of our homemade MUTATION standard compared to the Low MUTATION Standard using DHPLC analysis.