DNA replication sites

Camille Clément
Application specialist @Abbelight

 

COS7 cells incubated with EdU for 30 minutes.

Triton extraction and fixation.

EdU labeling with Alexa Fluor 647 using Click-Chemistry.

3D-dSTORM using DAISY technology

Clustering

DNA replication sites can be visualized using 5-ethynyl-2′-deoxyuridine (EdU). EdU is a thymidine analogue, which is incorporated into the DNA of replicating cells. Post fixation, it can be coupled with a fluorophore and visualized in microscopy. Incubation of cells with EdU for 30 minutes allows the visualization of DNA sites that replicated during the 30 minute-incubation. The combination of dSTORM and EdU labeling provides a powerful tool to study DNA replication with high precision.