CometAssay Single Cell Gel Electrophoresis Assay

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed 1–2 × 10^4 cells/ml		Corresponding TIFF files were processed using the ImageJ plugin Open Comet (http://www.cometbio.org/), and EdU+ cells were manually scored for each cell ID generated.

Upstream tips
Seed 1–2 × 10^4 cells/ml
Downstream tips
Corresponding TIFF files were processed using the ImageJ plugin Open Comet (http://www.cometbio.org/), and EdU+ cells were manually scored for each cell ID generated.

Publication protocol

The neutral comet assay was performed according to manufacturer protocol (Trevigen #4250-050-K) with an adaptation to capture EdU staining. Briefly, after treating cells for 30 min with EdU, cell suspensions were harvested and suspended in LM agarose gels on comet slides and processed for single-cell electrophoresis and DNA precipitation. To stain, gel sites were immersed in EdU click reaction cocktail with Alexa Fluor 647 for 30 min at room temperature. Slides were washed and then stained with SYBR Gold (ThermoFisher, #S11494) for 30 min at room temperature. Slides were washed and dried thoroughly for 45 min before applying glycerol mountant and coverslips. Cometimages were taken on a Nikon Ti-E using a 10×0.45 NA objective. CorrespondingTIFFfiles were processed using the ImageJ plugin OpenComet (http://www.cometbio.org/), and EdU+cells were manually scored for each cell ID generated

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Manufacturer protocol

Download the product protocol from Bio-Techne for CometAssay Single Cell Gel Electrophoresis Assay below.

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