Characterization of aberrant melting peaks in unlabeled probe assays.

  title={Characterization of aberrant melting peaks in unlabeled probe assays.},
  author={Shale A Dames and Rebecca L. Margraf and David Pattison and Carl T. Wittwer and Karl V. Voelkerding},
  journal={The Journal of molecular diagnostics : JMD},
  volume={9 3},
An unlabeled probe assay relies on a double-stranded DNA-binding dye to detect and verify target based on amplicon and probe melting. During the development and application of unlabeled probe assays, aberrant melting peaks are sometimes observed that may interfere with assay interpretation. In this report, we investigated the origin of aberrant melting profiles observed in an unlabeled probe assay for exon 10 of the RET gene. It was determined that incomplete 3' blocking of the unlabeled probe… CONTINUE READING