Microarray analysis of DNA replication timing.
Although all of the DNA in an eukaryotic cell replicates during the S-phase of cell cycle, there is a significant difference in the actual time in S-phase when a given chromosomal segment replicates. Methods are described here for generation of high-resolution temporal maps of DNA replication in synchronized human cells. This method does not require amplification of DNA before microarray hybridization and so avoids errors introduced during PCR. A major advantage of using this procedure is that it facilitates finer dissection of replication time in S-phase. Also, it helps delineate chromosomal regions that undergo biallelic or asynchronous replication, which otherwise are difficult to detect at a genome-wide scale by existing methods. The continuous TR50 (time of completion of 50% replication) maps of replication across chromosomal segments identify regions that undergo acute transitions in replication timing. These transition zones can play a significant role in identifying insulators that separate chromosomal domains with different chromatin modifications.
Karnani
Neerja
N
Department of Biochemistry and Molecular Genetics, University of Virginia, Charlottesville, VA, USA.
Taylor
Christopher M
CM
Dutta
Anindya
A
D000465:Algorithms; D042522:DNA Replication Timing / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D046228:Microarray Analysis / Q000295:instrumentation / Q000379:methods*; D016196:S Phase / Q000502:physiology; D013997:Time Factors