Single tube optimisation of APOE genotyping based on melting curve analysis

Objectives To develop and validate a single-tube protocol for APOE genotyping using fluorescent probes. Design and methods We have designed and validated a hybrid, single-tube, SimpleProbe/FRET probe protocol that takes advantage of the different probe wavelength emissions. Results Our method offers high quality outcomes, minimum interferences between probe signals and is 100% concordant with the reference protocol. Conclusions This method is cheaper, faster and more reliable and versatile than other alternatives proposed.