Does anyone use the MS-HRM method to determine methylation in a given locus of DNA? What software do you use to design primers for this method? The most recommended is Methyl Primer Express (Thermofisher). After importing the DNA sequence I used the primer design function for Bisulfite Sequencing PCR (BSP). However, the designed primers had in their sequence degenerate nucleotides which is not advisable in MS-HRM. Have any of you used Methyl Primer Express to design primers for MS-HRM and know how to configure this software to design such primers? Do you use the Bisulfite Sequencing PCR (BSP) or methylation-specific PCR (MSP) option in this program? Are degenerate nucleotides in primers for MS-HRM inadvisable? I would appreciate any tips on designing such primers because I am stuck and don't know what software to use and how to configure it to get good primers for MS-HRM.
- Badges
- Science method