Good primer design tips
1. Primers should be 17-24 bases in length
2. Base composition should be 45-65% (G+C)
3. Primers should end (3′) in a G or C, or CG or GC: this prevents “breathing” of ends and increases efficiency of priming
4. Tms between 55-75ºC are preferred
5. Primer self-complementarity (ability to form 2º structures such as hairpins or primer dimers) [...]