TABLE 2.
Oligonucleotide primers and probes and the number of target copies with the resulting Ct values from specificity experiments
| Primers and probes | nifH clone | Oligonucleotide | Sequence (5′-3′)a | No. of target copies | Ct ± SDb |
|---|---|---|---|---|---|
| 907h22 | 907h22 | Forward | ACGGCGGAACTTGGTGTGT | 1.7 × 105 | 21.98 ± 0.07 |
| Reverse | AATACCGCGACCTGCACAAC | ||||
| Probe | CGGTGGTCCTGAGCCGGGAGTT | ||||
| 911h7 | Forward | ATGGCGGAACCTGGTGTGT | 1.5 × 105 | 43.84 ± 2.01 | |
| Reverse | GATGCCACGACCGGCACAGC | ||||
| Probe | CGGAGGTCCGGAACCGGGGGTT | ||||
| 912h4 | 912h4 | Forward | GGTTATGGACAAGGTCCGTGAA | 1.6 × 105 | 22.04 ± 0.05 |
| Reverse | AGCCGCGTTTACAGACATCTTC | ||||
| Probe | AGTTCCAGATCCTCAACGGTGCCGA | ||||
| 914h6 | Forward | GGTCATGGACAAAGTCCGTGAC | 1.4 × 105 | 45.00 ± 0.00 | |
| Reverse | AGCCCCACTTCAACACATCTTC | ||||
| Probe | AGGTCCAGATCCTCGACGGTTCCGA | ||||
| 910h3 | Forward | GGTTATGGACAAGGTCCGTGAA | 1.6 × 105 | 45.00 ± NA | |
| Reverse | AACCCTTGCGACAAACATCTTC | ||||
| Probe | AACTCGAGGTCCTCAACGGTACCGA | ||||
| 911h10 | Forward | GGTTATGGACAAGGTCCGCGAA | 1.8 × 105 | 25.66 ± NA | |
| Reverse | AGCCGCGTTTACAGACATCTTC | ||||
| Probe | AGTTCCAGGTCCTCTACGGTGCCGA |
a
Mismatched bases are shown underlined in the negative control sequences.
b
Triplicate reactions; NA, replicate reactions were not conducted.