MINCE-Seq
MINCE-Seq
Nascent Chromatin with EdU and sequencing (MINCE-seq) was developed to characterize the genome-wide location of nucleosomes and other chromatin proteins behind replication forks at high temporal and spatial resolution
In MINCE-seq, newly replicated DNA is labeled with the nucleotide analog ethynyl deoxyuridine (EdU), which is coupled with biotin ex vivo using click chemistry. Coupling with biotin ensures highly specific purification of newly replicated DNA from asynchronous cells, even if it is only a fraction of a percentage of total DNA. Micrococcal nuclease (MNase) treatment recovers DNA fragments bound both by nucleosomes and by non-histone DNA-binding proteins, which enables the mapping of newly replicated chromatin at near base-pair resolution.