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. 2010 Jan 26;5(1):e8888. doi: 10.1371/journal.pone.0008888

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Huang et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) MS analysis of the nuclease P1 digestion products of the oligonucleotides used in Figure 2A , before (upper panel) and after (lower panel) bisulfite treatment. (B) To determine the conversion efficiency of 5-hmC to CMS in the oligonucleotide shown in Figure 2A , a standard curve was used to determine the unknown quantity of hmdCMP in the sample before and after treatment with sodium bisulfite (see text for details). The absolute value of the intercept of the best-fit line with the X-axis gives the concentration of hmdCMP remaining in the sample after bisulfite treatment as 4.69 nM. Given that the hmdCMP concentration before bisulfite treatment was 1.5 µM, this corresponds to a conversion efficiency as high as 99.7%.