No. | Orientation | Primer (5′-3′) | Score [%] | nt changes | Tm [°C] | nt | GC% | 3′ΔG | 5′ΔG | Runs | Rep. | RE sites | 3′ GC% | Comments |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
1 | Forward | ACCACTGTTGCATATGCTGATCTGCATCC | 69 | 3 | 72.16 | 29 | 48.28 | −8.07 | −7.71 | 0 | 0 | 1 | 50 | RE sites: NdeI @11 |
2 | Forward | ACCACTGTTGCATATGCTGATCTGCATCCT | 70 | 3 | 72.75 | 30 | 46.67 | −7.72 | −7.71 | 0 | 0 | 1 | 50 | RE sites: NdeI @11 |
3 | Forward | ACCACTGTTGCATATGCTGATCTGCATCCTG | 69 | 3 | 74.64 | 31 | 48.39 | −8.2 | −7.71 | 0 | 0 | 1 | 75 | RE sites: NdeI @11 |
4 | Forward | ACCACTGTTGCATATGCTGATCTGCATCCTGT | 64 | 3 | 75.06 | 32 | 46.88 | −7.96 | −7.71 | 0 | 0 | 1 | 50 | RE sites: NdeI @11 |
5 | Forward | ACCACTGTTGCATATGCTGATCTGCATCCTGTG | 66 | 3 | 76.82 | 33 | 48.48 | −6.85 | −7.71 | 0 | 0 | 1 | 50 | RE sites: NdeI @11 |
22 | Reverse | GCAGATCAGCATATGCAACAGTGGTTAGAGTAG | 73 | 3 | 70.8 | 33 | 45.45 | −5.5 | −8.27 | 0 | 0 | 1 | 50 | RE sites: NdeI @10 |
23 | Reverse | TGCAGATCAGCATATGCAACAGTGGTTAGAGTAG | 73 | 3 | 72.58 | 34 | 44.12 | −5.5 | −8.65 | 0 | 0 | 1 | 50 | RE sites: NdeI @11 |
24 | Reverse | ATGCAGATCAGCATATGCAACAGTGGTTAGAGTAG | 73 | 3 | 72.64 | 35 | 42.86 | −5.5 | −8.52 | 0 | 0 | 1 | 50 | RE sites: NdeI @12 |
25 | Reverse | GCAGATCAGCATATGCAACAGTGGTTAGAGTA | 68 | 3 | 70.2 | 32 | 43.75 | −5.48 | −8.27 | 0 | 0 | 1 | 25 | RE sites: NdeI @10 |
26 | Reverse | TGCAGATCAGCATATGCAACAGTGGTTAGAGTA | 68 | 3 | 72.04 | 33 | 42.42 | −5.48 | −8.65 | 0 | 0 | 1 | 25 | RE sites: NdeI @11 |