Table 1 The primers for site-directed mutagenesis
From: A mutant screening method by critical annealing temperature-PCR for site-directed mutagenesis
Primer | Sequence | Length | Mismatches | T m |
|---|---|---|---|---|
D112G(F) | CTATGgCCTCCTAAAGGACCTAGAGGAAG | 29 | 1 | 76.0 |
D112G(R) | GGAGGcCATAGACGTTGCTGTCAGAG | 26 | 1 | 75.3 |
D112K(F) | CTATaAaCTCCTAAAGGACCTAGAGGAAG | 29 | 2 | 68.3 |
D112K(R) | GGAGtTtATAGACGTTGCTGTCAGAG | 26 | 2 | 66.8 |
I4V(F) | CAACCgTTCCCTTATCCAGGCTTTTTG | 27 | 1 | 72.5 |
I4V(R) | GAAcGGTTGGGAACATATGTATATCTCCTTCT | 32 | 1 | 73.9 |
L45D(F) | CATTCgatCAGAACCCCCAGACCTCCCTCTGTTTCT | 36 | 3 | 76.1 |
L45D(R) | GGTTCTGatcGAATGAATACTTCTGTTCCTTTGGGAT | 37 | 3 | 71.8 |
E56D(F) | TCAGAcTCTATTCCGACACCCTCCAAC | 27 | 1 | 74.1 |
E56D(R) | GAATAGAgTCTGAGAAACAGAGGGAGGTCTG | 31 | 1 | 76.3 |
R64D(F) | CTCCAACgacGAGGAAACACAACAG | 25 | 3 | 63.8 |
R64D(R) | CCTCgtcGTTGGAGGGTGTCGGAATAG | 27 | 3 | 69.7 |
N109Y(F) | CAGCtAtGTCTATGACCTCCTAAAGGACCTAGAGGAAG | 38 | 2 | 77.9 |
N109Y(R) | GACaTaGCTGTCAGAGGCGCCGTACACCAG | 30 | 2 | 76.9 |
D116F(F) | AAAGttCCTAGAGGAAGGCATCCAAACGCTGAT | 33 | 2 | 73.6 |
D116F(R) | CTCTAGGaaCTTTAGGAGGTCATAGACGTTGCTGTCAG | 38 | 2 | 77.9 |
R64M(F) | TCCAACAtGGAGGAAACACAACAG | 24 | 1 | 68.0 |
R64M(R) | TCCaTGTTGGAGGGTGTCGGAATAG | 25 | 1 | 71.8 |
hGH2V3-P233S(F) | TGGtCgTGGAAGgTGCTACTCCAGTGCCCACCAGCC | 36 | 3 | 80.6 |
hGH2V3-P233S(R) | GAGTAGCAcCTTCCAcGaCCAGGAGAGGCACTGGG | 35 | 3 | 79.4 |
hGHBP-S237C(F) | GATGtGCTAAGAATTCGAGCTCCGTCGACAA | 31 | 1 | 76.3 |
hGHBP-S237C(R) | TTAGCaCATCTGAGGAAGTGTTACATAGAGCACC | 34 | 1 | 76.8 |
pGHBP-S237C(F) | GATGtGCTAAGAATTCGAGCTCCGTCGACAA | 31 | 1 | 76.3 |
pGHBP-S237C(R) | TTAGCaCATCTGAGGAAGTGTTACATAGAGCACC | 34 | 1 | 76.8 |
hGHR1-255R235C(F) | AACAAtgcAACTCTGGAAATTATGGC | 26 | 3 | 59.8 |
hGHR1-255R235C(R) | TTgcaTTGTTTGGATCTCACA | 21 | 3 | 58.7 |
hGHR1-638R235C(F) | AACAAtgcAACTCTGGAAATTATGGC | 26 | 3 | 59.8 |
hGHR1-638R235C(R) | TTgcaTTGTTTGGATCTCACA | 21 | 3 | 58.7 |