Table 1.

Selected parameter settings for round one in the primer design process.

Primer length	18 – 22 nucleotides
Product length	80 – 120 nucleotides
Primer melting temperature	58.0 – 60.0 °
Max. primer melting temperature diff.	2.0 °
Enforce G/C terminal	Yes
G/C content	40.0 – 60.0%
Max. base repeat	2 (e.g., GGG in a primer is disallowed)
Force specificity for both primers	Yes (i.e., none of the two primers has a perfect alignment to a non-target)