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Figure 2 | Biological Procedures Online

Figure 2

From: A Modified Protocol for Bisulfite Genomic Sequencing of Difficult Samples

Figure 2

Rates of nucleotide conversion using the original or the modified protocols. The conversion status of 74 non-CpG-cytosine residues of the RARB2 P2 promoter was analyzed following conversion using the standard BGS protocol (blue bars) or the MRED modification (red bars). Clones were sequenced and grouped according to the percentage of converted cytosine residues. Results show an increase in the frequency of cytosine to uracil conversion using our modified protocol. While only 10% of samples sequenced using the original protocol (n = 200) achieved 99% conversion of non-CpG-cytosines, 91% of samples sequenced using the modified protocol (n = 176) achieved 99% conversion. The threshold for the status of informativity was randomly set to 73/74 (99%) conversions to uracil, and this threshold was used to distinguish fully converted sequencings from partially converted ones.

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Biological Procedures Online

ISSN: 1480-9222

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