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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