A number of reviews have accompanied and monitored the progress of molecular phylogenetic research on bryophytes, focusing on the publication record itself, bryophyte phylogeny and systematics in the molecular era, as well as the evolution and phylogenetic utility of markers from different genomes. However, none of the recent reviews include a detailed characterization of all molecular markers used in bryophyte phylogenetics. Here we provide an overview of the history and current state of marker utilization, including coding and non-coding sequence markers from all three genomes as well as fingerprinting approaches. The molecular architecture and evolutionary peculiarities, as well as practical aspects such as amplification and sequencing strategies, are outlined for the DNA sequence markers, with a focus on the most commonly employed regions. Their phylogenetic utility and potential for solving some of the remaining, pressing questions in bryophyte phylogeny, as well as their suitability for molecular species identification by DNA barcoding, are discussed.