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. Overall design: 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).