How should I design a primer for DNA sequencing Sample requirements for primers The melting temperature of the primer should be between 52° and 58°C and the length should be between 17 – 19bp. Ideally, the GC content of an 17mer should be 10 G+C; for an 18mer 8 – 9 G+C and for a 19mer 7 – 9 G+C. G or C should be at the 3’ end, but not more than 3 Gs or Cs. The primer sequence should be a good mix of all 4 nucleotides with no more than 4 identical bases in a row (AAAA or GGGG).
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