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Multiplexed TrAEL-seq captures DNA replication dynamics in mammalian cells - PubMed

3 hours ago
  • #genome profiling
  • #mammalian cells
  • #DNA replication
  • TrAEL-seq is a method for profiling DNA replication genome-wide in unsynchronized cells without drugs or nucleotide analogues.
  • An updated TrAEL-seq protocol improves sample quality and allows multiplexing of up to six samples, enhancing throughput.
  • The method provides high-resolution replication profiles comparable to OK-seq in mammalian cell lines.
  • TrAEL-seq is effective in dynamic systems, such as during oncogene-induced senescence.
  • It maps replication initiation and termination zones and is sensitive to replication fork speed.
  • Replication forks slow down in transcribed regions but this does not broadly affect replication dynamics.
  • Forks accelerate over the first ~1 Mb of travel, regardless of local transcription.
  • Fewer active forks later in S-phase and less competition for replication factors may explain this acceleration.