Estimating TimeTrees with MEGA and the TimeTree Resource

The main outcome of molecular dating, the timetree, provides crucial information for understanding the evolutionary history of lineages and is a requirement of several evolutionary analyses. Although essential, the estimation of divergence times from molecular data is frequently regarded as a complicated task. However, establishing biological timescales can be performed in a straightforward manner, even with large, genome-wide data sets. This protocol presents all the necessary steps to estimate a timetree in the program MEGA X. It also illustrates how the TimeTree resource can be a useful tool to obtain chronological information based on previous studies, therefore yielding calibration boundaries.