How should I prepare my primers?

If you are providing your own primers, please follow the following recommendations:

  1. Provide at least 5 uM of primer per reaction. (at a concentration between 5-10 µM)
  2. Select and design a primer that only primes at one location on your template and does not form secondary structures or primer dimers.
  3. The ideal primer length for sequencing is typically between 18 and 24 bp.
  4. Ideal Tm should be 60° C, as we can not modify the annealing temperature on our PCR machines for individual orders.
  5. Primers should have about 50% G-C content.