Cells were exposed at 0.5 μmol/L AZD6738, 0.5 μmol/L cisplatin, or their combination for 5 days. The cells were harvested, resuspended at 1×105 cells/mL in ice-cold phosphate-buffered saline (PBS), and combined with molten LMA-garose at a ratio of 1:10. Next, the samples were slightly moved onto comet slides and incubated at 4°C in the dark for 40 minutes. After immersion in precooled lysis solution at 4°C for 40 minutes, the slides were placed in freshly prepared alkaline unwinding solution (200 mM NaOH, 1 mM EDTA, pH > 13) for 30 minutes at room temperature in the dark. Electrophoresis was conducted for 30 minutes, and the samples were dried at room temperature overnight. Next, 100 μL of diluted SYBR Green staining solution was placed onto each circle of agarose, and the samples were covered with a coverslip. Tail moment and intensity were measured using the Comet Assay IV program (Andor Technology, Belfast, UK). Three independent experiments were analyzed for each condition.
Comet assay 4 program
Manufactured by Oxford Instruments
Sourced in United Kingdom
The Comet assay IV program is a software package designed for the analysis and quantification of DNA damage in individual cells. It provides a comprehensive set of tools for the automated scoring and interpretation of comet assay images, a widely used technique in genotoxicity testing and DNA damage assessment.
Automatically generated - may contain errors
5 protocols using comet assay 4 program
1
Comet Assay for DNA Damage Assessment
2
Alkaline Comet Assay for DNA Damage
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Alkaline comet assays were conducted using a Trevigen Comet Assay kit (Trevigen, Gaithersburg, MD, USA) according to the manufacturer’s instructions. Incorporated sybr-green was detected using a Zeiss LSM 800 laser scanning microscope. Photographs were taken at a magnification of X20, and tail intensities were measured using the Comet assay IV program (Andor Technology, Belfast, UK).
3
Alkaline Comet Assay Protocol
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An alkaline comet assay using a Trevigen Comet assay kit (Trevigen, Gaithersburg, MD, USA) was performed following the manufacturer’s protocol. Tail lengths were measured with the Comet assay IV program (Andor technology, Belfast, UK).
4
Comet Assay to Assess DNA Damage
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Cells were exposed to 1 μmol/L AZD1775, 1 μmol/L AZD6738, or both for 24 hours. Then, the cells were harvested, and alkaline comet assay was performed as previously described [11 (link)]. The tail moment and intensity were measured using the Comet Assay IV program (Andor Technology, Belfast, UK). Three independent experiments were analyzed for each condition.
5
Comet Assay for DNA Damage
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Comet assay was performed according to the manufacturer’s instructions (Trevigen), tail intensity was measured for 50 cells/condition with the Comet assay IV program (Andor Technology). Experiments were repeated independently three times.
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