Figure 3.

DNA immunoprecipitation with anti-5mC antibody. Immunoprecipitation with an antibody against 5-methylcytidine was carried out to test the antibody’s affinity towards modified cytosines. The levels of immunocaptured 76-mers were measured using liquid scintillation counting. As a control, normal mouse IgG was used for immunoprecipitation. The oligonucleotides C76, 5mC76 and 5hmC76 were synthesized by PCR and contain C, 5mC or 5hmC at three BstUI sites as schematically indicated at the top of the Figure (see Figure 1 for sequence). Oligonucleotides 5mC5hmC76 and 5mC76a were prepared by chemical synthesis and contain 5hmC or C at the central BstUI site. Experiments were done in triplicates and the standard deviation is shown.