Beside being able to use EasyBeacons™ in real-time measurements, they can also be used for end-point detection and/or verification.

This is an valuable feature especially if you want to detect or identify a genetic sequence or if you want quality control on your quantification assay.

As the name end-point detection indicates, the measurement is done after the amplification is finished.

A typical protocol for end-point detection is shown below.

The end point verification is carried out after the amplification has finished.
The temperature are increased to denaturation and then lowered to a starting temperature that is above the probes affinity. Then increased to denaturation again and lowered at a temperature lower than the previous measurement, this step is continued until the affinity temperature is determined. At the end of each step the fluorescence of the labels should be measured.

The fluorescence will increase when the EasyBeacon™ binds, and the affinity temperature is found as the first derivative of the curve.

The Figure below shows data from an end-point verifiacation curve from a study on detection of single CpG duplets.