Table 1.
Genes/loci used in the study with PCR primers, references and protocols.
| Locus a | Primers b | PCR: Thermal Cycles: c (Annealing temp. in Bold) |
References |
|---|---|---|---|
| act | ACT-512F ACT2Rd |
(96 °C: 120 s, 52 °C: 60 s, 72 °C: 90 s) × 40 cycles | [41,42] |
| btub | TUB2Fw TUB4Rd |
(94 °C: 30 s, 56 °C: 45 s, 72 °C: 60 s) × 35 cycles | [43] |
| cal | CAL-235F CAL2Rd |
(96 °C: 120 s, 50 °C: 60 s, 72 °C: 90 s) × 40 cycles | [42,44] |
| ITS | ITS5 ITS4 |
(95 °C: 30 s, 55 °C:50 s, 72 °C: 90 s) × 35 cycles | [45] |
| LSU | LR0R LR5 |
(95 °C: 30 s, 55 °C:50 s, 72 °C: 90 s) × 35 cycles | [46,47] |
| rpb2 | fRPB2-5f fRPB2-7cR |
(94 °C: 60 s, 58 °C: 60 s, 72 °C: 90 s) × 40 cycles | [48] |
| fRPB2-414R | (96 °C: 120 s, 49 °C: 60 s, 72 °C: 90 s) × 40 cycles | [49] | |
| SSU | NS1 NS4 |
(95 °C: 30 s, 55 °C:50 s, 72 °C: 90 s) × 35 cycles | [45] |
| tef1 | EF1-983F EF1-2218R |
(95 °C: 30 s, 55 °C:50 s, 72 °C: 90 s) × 35 cycles | [50,51] |
| EF1-728F EF-2 |
(96 °C: 120 s, 52 °C: 60 s, 72 °C: 90 s) × 40 cycles | [41,52] |
aact: actin; btub: β-tubulin; cal: calmodulin; ITS: part of rDNA 18S (3’ end), the first internal transcribed spacer (ITS1), the 5.8S rRNA gene, the second ITS region (ITS2), and part of the 28S rRNA (5’ end); LSU: large subunit (28S); rpb2: RNA polymerase II second largest subunit; SSU: small subunit rDNA (18S); tef1: translation elongation factor 1-alpha gene. b fRPB2-5f and fRPB2-414R were used only for Teratosphaeriaceae analysis. c All the PCR thermal cycles include initiation step of 95 °C: 5 min, and final elongation step of 72 °C: 10 min and final hold at 4 °C.