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. 2002 Feb;22(3):704–723. doi: 10.1128/MCB.22.3.704-723.2002

FIG. 6.

FIG. 6.

Sodium bisulfite genomic sequencing of EBNA1 regions 2 and 6 from in vitro-methylated p220.2 DNA. (A) EBNA1 region 1 DNA (approximately 250 bp in length) with 11 CpG sites was PCR amplified, cloned, and sequenced after sodium bisulfite treatment of p220.2 DNA methylated by GST-3a in vitro. CpG site 3 is an HhaI site. (B) EBNA1 region 2 DNA (approximately 250 bp long) with 10 CpG sites from in vitro Dnmt3a methylated p220.2 was analyzed by the sodium bisulfite genomic sequencing method, as described above. CpG site 5 is an HhaI site. Plasmid DNA methylated and sequenced in independent experiments showed similar results. The filled circles represent methylated CpG sites, and the open circles represent unmethylated CpG sites. Plasmid DNA methylated and sequenced in independent experiments showed similar results. The frequency of methylation at each site and the overall methylation frequency of each region were calculated as described in the legend to Fig. 3.