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We carried out a melting analysis based on the principle of turbidity.
The allele specific amplicons were identified with high resolution melting analysis (HRM).
High-resolution melting analysis has been validated for screening several genes of clinical significance.
These results were confirmed by direct DNA sequencing and by high-resolution melting analysis.
High resolution melting analysis (HRMA) is a highly sensitive closed-tube genotyping method used primarily in clinical studies.
At the end of the PCR, samples were subjected to a melting analysis to confirm specificity of the amplicon.
Two different methods were used to genotype these SNPs: restriction fragment length polymorphisms or unlabelled probe melting analysis [37].
However, instruments vary widely in their ability to genotype variants by whole amplicon melting analysis [18], [19].
This off-line protocol permitted direct comparison of the melting analysis results from the same reaction on different platforms.
The time spent on melting analysis after PCR ranged from 15 min to 45 min. The absolute Tm values also varied in some genotype (within ±2°C).
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High-resolution melting analysis-identified mutations in 4/50 patients that had been missed by direct sequencing.
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