Skip to main content
Figure 3 | BMC Research Notes

Figure 3

From: Affinity-based enrichment strategies to assay methyl-CpG binding activity and DNA methylation in early Xenopus embryos

Figure 3

Genomic DNA isolation and sonication. (A) Xenopus tropicalis genomic DNA isolated from blastula (bl) and gastrula (ga) embryos. (B) Genomic DNA analyzed after 5 minutes or 15 minutes of sonication. Genomic DNA sonicated for 15 minutes yields fragments of 100 - 300 bp. (C) A schematic representation of methylated DNA affinity capture (MethylCap). Methylated DNA is bound by the immobilized methyl-CpG binding domain of MeCP2, washed and eluted with increasing NaCl concentrations. (D) PCR quantitation of methylated DNA recovery. DNA methylation targets (tcea, tfcp2 and gs17) mostly elute at high NaCl concentrations (500 mM, 600 mM and 700 mM) whereas the non-methylated targets (Xbra, trim33, hes4) are washed out with the first washing step (200 mM). (E) The methylation status of genomic targets tcea, tfcp2, gs17, trim33 and hes4 was validated by MeDIP (methylated DNA immunoprecipitation). The error bars represent standard deviation (SD) of two experiments.

Back to article page