Figure 8

Detection of methylation using methylation sensitive restriction enzyme cutting combined with PCR amplification. Chromosomal DNA of wild type and of AC2 expressing tobacco leaves were isolated and cut with either BamHI (1) or BamHI and HpaII (2) restriction enzymes. DNA fragments were amplified using primers presented in Additional file 11. Osmotin, AP-24 (A) and ethylene responsive factor1 ERF1 (B) indicated methylation in AC2 DNA fragments, whereas no methylation was detected in 18S rRNA (C) AC2 DNA fragments compared to wild type DNA samples. Data consists of four biological replicates.