Molecular combing solutions to characterize replication kinetics and genome rearrangements

Abstract Analyses of the DNA fibers have become a central technique to investigate the phenomena causing genomic instability in the field of cytogenomics. Molecular combing has achieved a high value in replication analyses and genome integrity with greater resolution and robustness. In the replication assay, cells are pulsed with two different thymidine analogs. While for the Genome Morse Code (GMC), specific probes are designed to build a unique pattern of color for the characterization of specific genome sequences. DNA molecules are combed at a specific parameter (2 kb per micrometer) on specific chemical-treated coverslips. Thus, DNA single molecules are parallel aligned with a constant stretching factor to each other. Molecular combing has strong value and potentiality for investigations regarding the replication stress response: such as monitoring changes in the kinetics of replication forks and origins. Alternatively, the GMC analyses bring to the identification and quantification of genomic rearrangements and genetic modifications.