Table 1.
Individual components and complete sequences of Illumina MiSeq-compatible custom sequencing primers used to sequence amplicon libraries in this study and their estimated melting temperatures.
| PCR direction and primers | Primer sequences (5′–3′) | Tm (°C) |
| Forward | ||
| 5′ Illumina adapter P5 | AATGATACGGCGACCACCGAGATCTACAC | — |
| Forward primer pad | TATGGTAATT | — |
| Forward primer linker | CT | — |
| Forward primer (NS31) | TTGGAGGGCAAGTCTGGTGCC | — |
| Complete forward primer construct (NS31f_il) | AATGATACGGCGACCACCGAGATCTACACTATGGTAATTCTTTGGAGGGCAAGTCTGGTGCC | 70.6 |
| Reverse | ||
| Reverse complement of 3′ Illumina adapter P7 | CAAGCAGAAGACGGCATACGAGAT | — |
| Golay barcode (see Appendix 2) | XXXXXXXXXXXX | — |
| Reverse primer pad | AGTCAGTCAG | — |
| Reverse primer linker | AC | — |
| Reverse primer (AML2) | GAACCCAAACACTTTGGTTTCC | — |
| Complete reverse primer construct (AML2r-il) | CAAGCAGAAGACGGCATACGAGATXXXXXXXXXXXXAGTCAGTCAGACGAACCCAAACACTTTGGTTTCC | 69.8–71.3 |
Note: Tm = melting temperature.