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Table 2 Features of the primers and their gradient-PCR identified- T c

From: A mutant screening method by critical annealing temperature-PCR for site-directed mutagenesis

Primer Length Mismatches %GC Nt3* Nt5Δ T m T c
D112G(F) 29 1 51.7 23 5 76.0 68
D112G(R) 26 1 57.7 20 5 75.3 68
D112K(F) 29 2 41.4 22 4 68.3 66
D112K(R) 26 2 46.2 19 4 66.8 66
I4V(F) 27 1 48.1 21 5 72.5 67
I4V(R) 32 1 40.6 28 3 73.9 67
L45D(F) 36 3 52.8 28 5 76.1 67
L45D(R) 37 3 40.5 27 7 71.8 67
E56D(F) 27 1 51.9 21 5 74.1 67
E56D(R) 31 1 48.4 23 7 76.3 67
R64D(F) 25 3 52.0 15 7 63.8 65
R64D(R) 27 3 59.3 20 4 69.7 65
N109Y(F) 38 2 47.4 31 4 77.9 69
N109Y(R) 30 2 60.0 24 3 76.9 69
D116F(F) 33 2 45.5 27 4 73.6 69
D116F(R) 38 2 52.0 29 7 77.9 69
R64M(F) 24 1 47.4 16 7 68.0 67
R64M(R) 25 1 40.5 21 3 71.8 67
hGH2V3-P233S(F) 36 3 63.9 19 3 80.6 70
hGH2V3-P233S(R) 35 3 62.9 17 4 79.4 70
hGHBP-S237C(F) 31 1 48.4 26 4 76.3 69
hGHBP-S237C(R) 34 1 44.1 28 5 76.8 69
pGHBP-S237C(F) 31 1 48.4 26 4 76.3 69
pGHBP-S237C(R) 34 1 44.1 28 5 76.8 69
hGHR1-255R235C(F) 26 3 38.5 18 5 59.8 60
hGHR1-255R235C(R) 21 3 38.1 16 2 58.7 60
hGHR1-255R235C(F) 26 3 38.5 18 5 59.8 60
hGHR1-255R235C(R) 21 3 38.1 16 2 58.7 60
  1. * The number of bases from 3- terminus to the mutation site.
  2. Δ The number of bases from 5-terminus to the mutation site.