Several things should be taken into consideration when designing methylation-specific primers for use with the EpiTect MSP Kit:
1. Each primer should include at least one but preferably two CpG sites.
2. Both primers should have CpG sites at or near their 3’ end.
3. Primers should contain cytosines which are not in a CpG site. This will make your primers
specific for bisulfite-converted DNA since all unmethylated cytosines will be converted to
uracils in bisulfite-treated DNA.
4. Primer pairs used for the detection of unmethylated and methylated DNA should cover the
same CpG sites. This is important to determine the methylation status of precisely defined
5. Amplicons should have a length between 100 and 200 base pairs.