RT Journal
A1 Debo, Brian M.
A1 Mallory, Benjamin J.
A1 Stergachis, Andrew B.
T1 Evaluation of N6-methyldeoxyadenosine antibody-based genomic profiling in eukaryotes
JF Genome Research 
JO Genome Research 
YR 2023 
FD March 01 
VO 33 
IS 3 
SP 427 
OP 434 
DO 10.1101/gr.276696.122 
UL http://genome.cshlp.org/content/33/3/427.abstract 
AB Low-level DNA N6-methyldeoxyadenosine (DNA-m6A) has recently been reported across various eukaryotes. Although anti-m6A antibody–based approaches are commonly used to measure DNA-m6A levels, this approach is known to be confounded by DNA secondary structures, RNA contamination, and bacterial contamination. To evaluate for these confounding features, we introduce an approach for systematically validating the selectivity of antibody-based DNA-m6A methods and use a highly selective anti-DNA-m6A antibody to reexamine patterns of DNA-m6A in C. reinhardtii, A. thaliana, and D. melanogaster. Our findings raise caution about the use of antibody-based methods for endogenous m6A quantification and mapping in eukaryotes.