IDT does not have a sequencing primer design service. We recommend using the PrimerQuest software available for free on our website to help you choose the primers.
The best sequencing data is found in the region 80-150bp away from the primer. It is difficult to get good base calls less than 50bp away from the primer and greater than 300bp away.
Length should be between 18 and 30 nt, with optimal being 20-25 nt.
G-C content of 40-60% is desirable.
The Tm should be between 55°C and 65°C.
Avoid primers with strong self-dimers or hairpins. You can use our OligoAnalyzer located on our website to check for this.
You also want to avoid primers which are >90% homologous to other regions within your sample. A BLAST search will alert you to this problem.