Methylation analysis of CDKN1C’s DMR in bovine . Restriction enzyme analysis was used to determine the methylation status of CDKN1C DMR in the bovine. The restriction enzymes HPAII (blocked by CpG methylation) and MSPI (able to digest both methylated and unmethylated CpGs) were used to determine the methylation of CDKN1C exons 1 through intron 2. M. Sss1 (methylates all CpGs) was used as a positive control to show that HPAII is unable to cleave methylated CpGs. Our results show that at least one of the 19 CCGG recognition sites for HPAII was unmethylated because there was no PCR amplification of this region for the HPAII digested template. H = Holstein, N = Nelore, F1 = B. t. indicus x B. t. taurus F1-C conceptus. - PCR = water PCR control to show no DNA contamination.