Genome-wide analysis of replication initiation in Tetrahymena [Vegetative growing cells]

We mapped Tetrahymena macronuclear replication origins on a genome-wide scale using nanopore sequencing. Nanopore sequencing quantifies individual BrdU incorporated replication intermediates on individual DNA molecules. The asynchronous exponentially growing Tetrahymena cells were pulse labeled with 100 µM (or 500 µM) of BrdU for 5 min and chased with 50 µM (or 250 µM) thymidine for 5 min, then added final 1 mM (or 2 mM) of thymidine for 20 min (Hennion et al. 2020).