Nature research paper: Genetic instability from a single S phase after whole-genome duplication
Comet assays were performed using Single Cell Gel Electrophoresis Assay kit according to the manufacturer’s instructions. Comets were then imaged using an inverted Eclipse Ti-E Nikon videomicroscope equipped with a 40× CFI Plan Fluor objective. Images were analysed with OpenComet plugin on Fiji. Based on the comet DNA content of DMSO treated cells, a manual threshold was applied to identify diploid from tetraploid cells.
Diploid controls and the tetraploid-enriched population were then pulse-labelled with 0.1 mM CIdU and 0.1 mM IdU for 30 min and 100,000–300,000 cells per condition were collected for further analysis. The DNA was extracted from cells and prepped following the manufacturer’s instructions using the FiberPrep DNA Extraction Kit . Subsequently, the prepped DNA was stretched onto coated glass coverslips by using the FiberComb Molecular Combing System .
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