Table 1. Molecular markers and primers used for amplification.
The amplification was performed in 50 µl final volume containing 18 µl of ultra-pure water (dd H2O), 25 µl of I-5TM 2X High-Fidelity Master Mix, 2 µl of each primer (100 pmol/ µl), 3 µl of the genomic spider DNA templates. PCR settings list Initial Denaturation (ind), followed by /n cycles (Denaturation: de, Primer Annealing: pra, Primer Elongation: pre), and one Terminal Elongation (tee). (Temperature in °C following by time in seconds).
| Marker | Primer name | Premier sequence (5′→ 3′) | PCR settings |
|---|---|---|---|
| 16S | 16SA | CGCCTGTTTACCAAAAACAT | ind98 (120s), [de 98(10s), pra 52(15s), pre72(15s)/35], tee72 (120s) |
| 16SB | CCGGTTTGAACTCAGATC | ||
| 18S | 18S5f | GCGAAAGCATTTGCCAAGAA | ind98 (120s), [de 98(10s), pra 57(15s), pre72(15s)/35], tee72 (120s) |
| 18S9r | GATCCTTCCGCAGGTTCACCTAC | ||
| 28S | 28SC | GGTTCGATTAGTCTTTCGCC | ind98 (120s), [de 98(10s), pra 55(15s), pre72(15s)/35], tee72 (120s) |
| 28SO | GAAACTGCTCAAAGGTAAACGG | ||
| COI | LCOI1490 | GGTCAACAAATCATAAAGATATTGG | ind98 (120s), [de 98(10s), pra 47(15s), pre72(15s)/35], tee72 (120s) |
| HCOI2198 | TAAACTTCAGGGTGACCAAAAAATCA | ||
| Jerry | CAACATTTATTTTGATTTTTTGG | ind98 (120s), [de 98(10s), pra 52(15s), pre72(15s)/35], tee72 (120s) | |
| C1-N-2776 | GGATAATCAGAATATCGTCGAGG | ||
| H3 | H3aF | ATGGCTCGTACCAAGCAGACVGC | ind98 (120s), [de 98(10s), pra 56(15s), pre72(15s)/35], tee72 (120s) |
| H3aR | ATATCCTTRGGCATRATRGTGAC | ||
| ITS2 | ITS4 | TCCTCCGCTTATTGATATGC | ind98 (120s), [de 98(10s), pra 50(15s), pre72(15s)/35], tee72 (120s) |
| ITS5.8 | GGGACGATGAAGAACGCAGC |