Table 1 Guidelines for choosing (MRED) restriction enzymes
From: A Modified Protocol for Bisulfite Genomic Sequencing of Difficult Samples
1 | The RE site must contain at least one non-CpG-cytosine |
|---|---|
2 | The RE site must be abolished following cytosine to uracil conversion |
3 | The most 5' and 3' nucleotides of the sequence must not form CpG dinucleotides with upstream or downstream sequences respectively |
4 | The RE site preferably does not have a site within the primer sequences |
5 | The RE site may be cut with a methylation-sensitive RE, since PCR products are not methylated |