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Measurement of Oxidatively Induced DNA Damage in Caenorhabditis elegans with High-Salt DNA Extraction and Isotope-Dilution Mass Spectrometry

Published

Author(s)

Leona D. Scanlan, Pawel Jaruga, Sanem Hosbas Coskun, Christopher M. Sims, Shannon Hanna, Jamie L. Almeida, David N. Catoe, Bryant C. Nelson, M Miral Dizdar, Erdem Coskun

Abstract

Caenorhabditis elegans (C. elegans) is used as a medical and systems toxicity model organism for environmental and developmental assays that include high-throughput methods and genetic studies. However, little is known about background levels of oxidatively induced DNA damage in the nematode or how culturing methods may affect DNA damage levels. Furthermore, C. elegans physiology makes it difficult to extract large amounts (> 50 µg) of high-quality genomic DNA. We therefore developed and optimized a DNA extraction method based on enzymatic digestion and high-salt phase separation that resulted in efficient extraction of > 50 µg DNA from 250000 to 500000 nematodes. Extracted DNA showed suitable RNA levels (
Citation
Analytical Chemistry
Created August 27, 2019, Updated October 8, 2019