The use of isolated peripheral lymphocytes and human whole blood in the comet assay

2024 · doi:10.21203/rs.3.nprot-5207/v2
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Abstract

Abstract The Comet assay, a highly sensitive method for detecting DNA damage, is used to measure DNA breaks and alkali-labile lesions in eukaryotic cells. This study outlines the application of whole blood in the alkaline gel electrophoresis method. A total of 1,200 blood samples from various individuals were analysed, comprising 300 samples from healthy individuals, 300 from suspected or pre-cancerous individuals, and 600 from cancer patients. Each sample was divided into two identical volumes in separate falcon tubes. The blood samples were prepared and preserved by adding an equal amount of RPMI medium and 10% DMSO. Analysis of the data using Student’s t-Test revealed a p-value of 0.59 for Olive tail moment (OTM) and 0.16 for % tail DNA, indicating no statistically significant differences between the two methods, whether with or without treatment. In conclusion, employing whole blood instead of isolated lymphocytes not only saves time but also maintains a high level of sensitivity, requiring less than 20 µL of blood from each individual.
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