Simplified method for the collection, storage, and comet assay analysis of DNA damage in whole blood

K. Al Salmani, H. H. Abbas, S. Schulpen, M. Karbaschi, I. Abdalla, K.J. Bowman, K. K. So, M. D. Evans, G.D. Jones, R.W.L. Godschalk, M. S. Cooke

Research output: Contribution to journalArticleAcademicpeer-review

16 Citations (Scopus)

Abstract

Single-cell gel electrophoresis (comet assay) is one of the most common methods used to measure oxidatively damaged DNA in peripheral blood mononuclear cells (PBMC), as a biomarker of oxidative stress in vivo. However, storage, extraction, and assay workup of blood samples are associated with a risk of artifactual formation of damage. Previous reports using this approach to study DNA damage in PBMC have, for the most part, required the isolation of PBMC before immediate analysis or freezing in cryopreservative. This is very time-consuming and a significant drain on human resources. Here, we report the successful storage of whole blood in ~250mul volumes, at -80 degrees C, without cryopreservative, for up to 1month without artifactual formation of DNA damage. Furthermore, this blood is amenable for direct use in both the alkaline and the enzyme-modified comet assay, without the need for prior isolation of PBMC. In contrast, storage of larger volumes (e.g., 5ml) of whole blood leads to an increase in damage with longer term storage even at -80 degrees C, unless a cryopreservative is present. Our "small volume" approach may be suitable for archived blood samples, facilitating analysis of biobanks when prior isolation of PBMC has not been performed.
Original languageEnglish
Pages (from-to)719-725
Number of pages7
JournalFree Radical Biology and Medicine
Volume51
Issue number3
DOIs
Publication statusPublished - 1 Jan 2011

Cite this

Al Salmani, K., Abbas, H. H., Schulpen, S., Karbaschi, M., Abdalla, I., Bowman, K. J., So, K. K., Evans, M. D., Jones, G. D., Godschalk, R. W. L., & Cooke, M. S. (2011). Simplified method for the collection, storage, and comet assay analysis of DNA damage in whole blood. Free Radical Biology and Medicine, 51(3), 719-725. https://doi.org/10.1016/j.freeradbiomed.2011.05.020