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. 2020 Mar 18;6(12):eaay3335. doi: 10.1126/sciadv.aay3335

Fig. 4. Artifactual detection of 6mdA in MS and single-molecule real-time sequencing.

Fig. 4

(A) Meta-analysis of published 6mdA abundance (% 6mdA/dA) in mammalian DNA as determined by high performance liquid chromatography–tandem MS for species human (n = 34), mouse (n = 18), pig (n = 11), and rat (n = 7) from nine independent studies. LOD, limit of detection. (B) Number of methylated CpGs (mCpGs) flanking (±12 bp) single-molecule real-time sequencing (SMRT-seq) 6mdA sites in human blood or matched random adenines (N = 881,240; equal chromosomal distribution). (C) mCpG distribution around SMRT-seq 6mdA sites in human blood or matched random adenines (N = 881,240; equal chromosomal distribution). (D) CpG distribution around SMRT-seq 6mdA sites in a human lymphoblastoid cell line (AK1) or matched random adenines (N = 80,560; equal chromosomal distribution). (E) Venn diagrams of SMRT-seq and DIP-seq peak overlaps for 6mdA in mouse (n = 4) and human (n = 4) samples. KO, knockout; PBMC, peripheral blood mononuclear cell.