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 |