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Table 3 Melting temperature (Tm) of each primer according to the number of mismatches allowed between the primer and the target sequence.

From: ITS as an environmental DNA barcode for fungi: an in silico approach reveals potential PCR biases

Primer Number of mismatches allowed
  0 1* 2* 3*
ITS1(1) ** 58.64 51.75+/-2.88 46.51+/-0.6 41.4+/-NA
ITS1(2) ** 58.64 52.02+/-2.58 46.46+/-0.87 39.49+/-2.75
ITS1-F 51.04 42.31+/-1.2 38.91+/-2.62 31.64+/-0.67
ITS2 56.68 48.5+/-1.97 39.3+/-2.74 32.99+/-5.67
ITS5 51.64 41.8+/-1.69 36.6+/-3.93 NA
ITS3 56.68 50.6+/-1.15 44.3+/-3.65 39.93+/-7.25
ITS4 50.9 45.04+/-1.3 35.94+/-3.38 32.73+/-1.83
ITS4-B 59.33 54.49+/-2.39 46.6+/-3.06 37.72+/-7.38
  1. * Mean Tm +/- SD is given for primers with 1 or more mismatches as the Tm depends on the type of mismatch.
  2. ** ITS1 is evaluated both with the first subset (1) and the second subset (2).