What are the unique features of the Ligase-65 enzyme?
Ligase-65 is active up to 65 °C, in contrast to for instance T4-Ligase. Contrary to Taq-Ligase and Tth-Ligase, the Ligase-65 enzyme is easily inactivated before the start of the PCR reaction by briefly heating the MLPA reactions to 95-98 °C. Ligase-65 is very specific in that it will only ligate perfectly matched probe-target hybrids. In comparison to other ligases tested for MLPA reactions at MRC-Holland, Ligase-65 resulted in the most reproducible results and the lowest number of aspecific bands.
