Measurement of Oxidatively Induced DNA Damage in <i>Caenorhabditis elegans</i> with High-Salt DNA Extraction and Isotope-Dilution Mass Spectrometry.

ABSTRACT: Caenorhabditis elegans is used extensively as a medical and toxicological model organism. However, little is known about background levels of oxidatively induced DNA damage in the nematode or how culturing methods affect DNA damage levels. The tough C. elegans cuticle makes it challenging to extract genomic DNA without harsh procedures that can artifactually increase DNA damage. Therefore, a mild extraction protocol based on enzymatic digestion of the C. elegans cuticle with high-salt phase-separation of DNA has been developed and optimized. This method allows for efficient extraction of >50 μg DNA using a minimum of 250000 nematodes grown in liquid culture. The extracted DNA exhibited acceptable RNA levels (<10% contamination), functionality in polymerase chain reacti

SUBMITTER: Scanlan LD 

PROVIDER: S-EPMC6996937 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature