Table 1.
Primer/probe set Name | Sequence(5′ → 3′) | The location on accession number | Product sizes (bp) | |
---|---|---|---|---|
F2 | ATGGAGCACCTGAGGCAGCACTGGACAACA | 3452–3481 | 221 | |
A1 | P1 | [FAM]CACTGGACAACACGAGCAACCCCACTGCTTA[dSpacer] TATAAAGCACCGTTCACA[C3-spacer] | 3470–3519 | 203 |
R1 | [Biotin]CGTTGAGAAGGGCACAGTCGTATTGAAACA | 3643–3672 | ||
F2 | ATGGAGCACCTGAGGCAGCACTGGACAACA | 3452–3481 | 258 | |
A2 | P1 | [FAM]CACTGGACAACACGAGCAACCCCACTGCTTA[dSpacer] TATAAAGCACCGTTCACA[C3-spacer] | 3470–3519 | 250 |
R2 | [Biotin]GCACGAGGAGTTCTTGGATCTCCGTGGCTC | 3680–3709 | ||
F2 | ATGGAGCACCTGAGGCAGCACTGGACAACA | 3452–3481 | 352 | |
A3 | P1 | [FAM]CACTGGACAACACGAGCAACCCCACTGCTTA[dSpacer] TATAAAGCACCGTTCACA[C3-spacer] | 3470–3519 | 229 |
R3 | [Biotin]GCAGGGGCAATAATTTTCTGCTTGTGTCTG | 3774–3803 | ||
F1 | CACCTGAGGCAGCACTGGACAACACGAGCAA | 3458–3488 | 215 | |
A4 | P1 | [FAM]CACTGGACAACACGAGCAACCCCACTGCTTA[dSpacer] TATAAAGCACCGTTCACA[C3-spacer] | 3470–3519 | 203 |
R1 | [Biotin]GCCGTAGTTGAAGGAGGCAGGAAGCTGTGC | 3643–3672 | ||
F1 | CACCTGAGGCAGCACTGGACAACACGAGCAA | 3458–3488 | 252 | |
A5 | P1 | [FAM] CACTGGACAACACGAGCAACCCCACTGCTTA[dSpacer] TATAAAGCACCGTTCACA[C3-spacer] | 3470–3519 | 240 |
R2 | [Biotin]GCACGAGGAGTTCTTGGATCTCCGTGGCTC | 3680–3709 | ||
A6 (Optimal set) | F1 | CACCTGAGGCAGCACTGGACAACACGAGCAA | 3458–3488 | 346 |
P1 | [FAM]CACTGGACAACACGAGCAACCCCACTGCTTA[dSpacer] TATAAAGCACCGTTCACA[C3-spacer] | 3470–3519 | 334 | |
R3 | [Biotin]GCAGGGGCAATAATTTTCTGCTTGTGTCTG | 3774–3803 | ||
F3 | TGAGGCAGCACTGGACAACACGAGCAACCC | 3462–3461 | 211 | |
A7 | P1 | [FAM]CACTGGACAACACGAGCAACCCCACTGCTTA[dSpacer] TATAAAGCACCGTTCACA[C3-spacer] | 3470–3519 | 203 |
R1 | [Biotin]GCCGTAGTTGAAGGAGGCAGGAAGCTGTGC | 3643–3672 | ||
F3 | TGAGGCAGCACTGGACAACACGAGCAACCC | 3462–3461 | 248 | |
A8 | P1 | [FAM]CACTGGACAACACGAGCAACCCCACTGCTTA[dSpacer] TATAAAGCACCGTTCACA[C3-spacer] | 3470–3519 | 240 |
R2 | [Biotin]GCACGAGGAGTTCTTGGATCTCCGTGGCTC | 3680–3709 | ||
F3 | TGAGGCAGCACTGGACAACACGAGCAACCC | 3462–3461 | 342 | |
A9 | P1 | [FAM]CACTGGACAACACGAGCAACCCCACTGCTTA[dSpacer] TATAAAGCACCGTTCACA[C3-spacer] | 3470–3519 | 334 |
R3 | [Biotin]GCAGGGGCAATAATTTTCTGCTTGTGTCTG | 3774–3803 | ||
F1 | AAAAGCAACCCATTACCCGCCTGGCACTCC | 3582–3611 | 285 | |
As1 | P1 | [FAM]ACCCGCCTGGCACTCCCTTACACCGCTCCC[dSpacer]ACCGTGTGCTTGCAACAGT[C3-spacer] | 3596–3645 | 271 |
R2 | [Biotin]GACTCTTCCCCGTAGGTTGTCTTCCCGTTG | 3837–3866 | ||
As2 | F1 | AAAAGCAACCCATTACCCGCCTGGCACTCC | 3582–3611 | 145 |
P1 | [FAM]ACCCGCCTGGCACTCCCTTACACCGCTCCC[dSpacer]ACCGTGTGCTTGCAACAGT[C3-spacer] | 3596–3645 | 131 | |
R3 | [Biotin]GGGAGTGCCAGGCGGGTAATGGGTTGCTTT | 3697–3726 | ||
F2 | AACCCAACCGCCTACCAAAAGCAACCCATT | 3566–3595 | 153 | |
As3 | P1 | [FAM]ACCCGCCTGGCACTCCCTTACACCGCTCCC[dSpacer]ACCGTGTGCTTGCAACAGT[C3-spacer] | 3596–3645 | 123 |
R1 | [Biotin]CGGTGTAAGGGAGTGCCAGGCGGGTAATGG | 3689–3718 | ||
F2 | AACCCAACCGCCTACCAAAAGCAACCCATT | 3566–3595 | 301 | |
As4 | P1 | [FAM]ACCCGCCTGGCACTCCCTTACACCGCTCCC[dSpacer]ACCGTGTGCTTGCAACAGT[C3-spacer] | 3596–3645 | 271 |
R2 | [Biotin]GACTCTTCCCCGTAGGTTGTCTTCCCGTTG | 3837–3866 | ||
F2 | AACCCAACCGCCTACCAAAAGCAACCCATT | 3566–3595 | 161 | |
As5 | P1 | [FAM]ACCCGCCTGGCACTCCCTTACACCGCTCCC[dSpacer]ACCGTGTGCTTGCAACAGT[C3-spacer] | 3596–3645 | 131 |
R1 | [Biotin]CGGTGTAAGGGAGTGCCAGGCGGGTAATGG | 3689–3718 | ||
F3 | CGAATCAGCAGACCCAGTTACCACCACAGT | 3274–3303 | 445 | |
As6 | P2 | [FAM]TGAAACTCACACAGCTCAAGAACACCCAAACT[dSpacer] TTGATCTTATGCAAATC[C3-spacer] | 3378–3427 | 341 |
R1 | [Biotin]CGGTGTAAGGGAGTGCCAGGCGGGTAATGG | 3689–3718 | ||
F3 | CGAATCAGCAGACCCAGTTACCACCACAGT | 3274–3303 | 593 | |
As7 | P2 | [FAM]TGAAACTCACACAGCTCAAGAACACCCAAACT[dSpacer] TTGATCTTATGCAAATC[C3-spacer] | 3378–3427 | 489 |
R2 | [Biotin]GACTCTTCCCCGTAGGTTGTCTTCCCGTTG | 3837–3866 | ||
F3 | CGAATCAGCAGACCCAGTTACCACCACAGT | 3274–3303 | 453 | |
As8 | P2 | [FAM]TGAAACTCACACAGCTCAAGAACACCCAAACT[dSpacer] TTGATCTTATGCAAATC[C3-spacer] | 3378–3427 | 349 |
R3 | [Biotin]GGGAGTGCCAGGCGGGTAATGGGTTGCTTT | 3697–3726 | ||
As9 (Optimal set) | F3 | CGAATCAGCAGACCCAGTTACCACCACAGT | 3274–3303 | 286 |
P2 | [FAM]TGAAACTCACACAGCTCAAGAACACCCAAACT[dSpacer] TTGATCTTATGCAAATC[C3-spacer] | 3378–3427 | 182 | |
R4 | [Biotin]GAGAAGTAGTACGTCGCAGACCGAAGTAGCG | 3530–3559 | ||
F3 | CGAATCAGCAGACCCAGTTACCACCACAGT | 3274–3303 | 445 | |
As10 | P1 | [FAM]ACCCGCCTGGCACTCCCTTACACCGCTCCC[dSpacer]ACCGTGTGCTTGCAACAGT[C3-spacer] | 3378–3427 | 341 |
R1 | [Biotin]CGGTGTAAGGGAGTGCCAGGCGGGTAATGG | 3689–3718 | ||
F3 | CGAATCAGCAGACCCAGTTACCACCACAGT | 3274–3303 | 593 | |
As11 | P1 | [FAM]ACCCGCCTGGCACTCCCTTACACCGCTCCC[dSpacer]ACCGTGTGCTTGCAACAGT[C3-spacer] | 3596–3645 | 271 |
R2 | [Biotin]GACTCTTCCCCGTAGGTTGTCTTCCCGTTG | 3837–3866 | ||
F1 | CAACACCACCAACCCAACGGCGTACCATAA | 3570–3599 | 161 | |
O1 | P1 | [FAM]CGTACCATAAGGCGCCGCTTACCCGGCTTA[dSpacer] ATTGCCCTACACGGCACCA[C3-spacer] | 3590–3639 | 141 |
R1 | [Biotin]GAGCCAGCACTTGGAGATCGCCTCTCACGT | 3701–3730 | ||
F1 | CAACACCACCAACCCAACGGCGTACCATAA | 3570–3599 | 343 | |
O2 | P1 | [FAM]CGTACCATAAGGCGCCGCTTACCCGGCTTA[dSpacer] ATTGCCCTACACGGCACCA[C3-spacer] | 3590–3639 | 323 |
R2 | [Biotin]CAAGGACTGCTTTACAGGTGCCACTATTTT | 3883–3912 | ||
F1 | CAACACCACCAACCCAACGGCGTACCATAA | 3570–3599 | 210 | |
O3 | P1 | [FAM]CGTACCATAAGGCGCCGCTTACCCGGCTTA[dSpacer] ATTGCCCTACACGGCACCA[C3-spacer] | 3590–3639 | 190 |
R3 | [Biotin]TTGATGGCACCGTAGTTGAAAGAAGTAGGC | 3751–3779 | ||
F2 | GGAGCACCTGAAGCAGCCTTGGACAACACC | 3549–3578 | 182 | |
O4 | P1 | [FAM]CGTACCATAAGGCGCCGCTTACCCGGCTTA[dSpacer] ATTGCCCTACACGGCACCA[C3-spacer] | 3590–3639 | 141 |
R1 | [Biotin]GAGCCAGCACTTGGAGATCGCCTCTCACGT | 3701–3730 | ||
F2 | GGAGCACCTGAAGCAGCCTTGGACAACACC | 3549–3578 | 364 | |
O5 | P1 | [FAM]CGTACCATAAGGCGCCGCTTACCCGGCTTA[dSpacer] ATTGCCCTACACGGCACCA[C3-spacer] | 3590–3639 | 323 |
R2 | [Biotin]CAAGGACTGCTTTACAGGTGCCACTATTTT | 3883–3912 | ||
O6 (Optimal set) | F2 | GGAGCACCTGAAGCAGCCTTGGACAACACC | 3549–3578 | 231 |
P1 | [FAM]CGTACCATAAGGCGCCGCTTACCCGGCTTA[dSpacer] ATTGCCCTACACGGCACCA[C3-spacer] | 3590–3639 | 190 | |
R3 | [Biotin]TTGATGGCACCGTAGTTGAAAGAAGTAGGC | 3751–3779 | ||
F3 | GGGGACCTTACCTGGGTGCCAAATGGAGCA | 3524–3553 | 217 | |
O7 | P2 | [FAM]CAAATGGAGCACCTGAAGCAGCCTTGGACAA[dSpacer]ACCACCAACCCAACGGCGTAC[C3-spacer] | 3542–3596 | 189 |
R1 | [Biotin]GAGCCAGCACTTGGAGATCGCCTCTCACGT | 3701–3730 | ||
F3 | GGGGACCTTACCTGGGTGCCAAATGGAGCA | 3524–3553 | 389 | |
O8 | P2 | [FAM]CAAATGGAGCACCTGAAGCAGCCTTGGACAA[dSpacer]ACCACCAACCCAACGGCGTAC[C3-spacer] | 3542–3596 | 371 |
R2 | [Biotin]CAAGGACTGCTTTACAGGTGCCACTATTTT | 3883–3912 | ||
F3 | GGGGACCTTACCTGGGTGCCAAATGGAGCA | 3524–3553 | 256 | |
O9 | P2 | [FAM]CAAATGGAGCACCTGAAGCAGCCTTGGACAA[dSpacer]ACCACCAACCCAACGGCGTAC[C3-spacer] | 3542–3596 | 238 |
R3 | [Biotin]TTGATGGCACCGTAGTTGAAAGAAGTAGGC | 3751–3779 |
Note: F:forward primer, R reverse primer, P probe, FAM Carboxyfluorescein, dSpacer A tetrahydrofuran residue, C3-spacer 3’-block