Table 1. PCR primers and conditions applied for molecular fungal identification.
| Locus | Amplicon length | Primer | Primer sequence 5’-3’ | PCR conditions | Reference |
|---|---|---|---|---|---|
| Internal Transcribed Spacer (ITS) region of the rRNA | ~450-800bp | ITS1F | CTT GGT CAT TTA GAG GAA GTA A | 1. 95˚C– 5 min 2. 95˚C– 30 s 3. 52˚C– 30 s 4. 72˚C– 1 min Steps 2–4 x 35 cycles 5. 72˚C– 8 min |
[18] |
| ITS4 | TCC TCC GCT TAT TGA TAT GC | ||||
| Translation elongation factor 1-alpha (tef1) | ~600 bp | EF1-1018F | GAY TTC ATC AAG AAC ATG AT | 1. 95˚C– 2 min 2. 66˚C-56˚C touchdown (9 cycles) 3. 95˚C– 30 s 4. 56˚C– 1 min 5. 72˚C– 1 min Steps 3–5 x 36 cycles 6. 72˚C– 10 min |
[19] |
| EF1-1620R | GAC GTT GAA DCC RAC RTT GTC | ||||
| beta-tubulin (tub2) | ~ 500 bp | Bt2a | GGT AAC CAA ATC GGT GCT GCT TTC | 1. 95˚C– 3 min 2. 95˚C– 30 s 3. 58˚C– 30 s 4. 72˚C– 1 min Steps 2–4 x 35 cycles 5. 72˚C– 10 min |
[20] |
| Bt2b | ACC CTC AGT GTA GTG ACC CTT GGC | ||||
| Translation elongation factor 1-alpha (tef1) | ~700 bp | ef1 | ATG GGT AAG GA(A/G) GAC AAG AC | 1. 95˚C– 3 min 2. 95˚C– 30 s 3. 58˚C– 30s 4. 72˚C– 1 min Steps 2–4 x 35 cycles 5. 72˚C– 10 min |
[21] |
| ef2 | GGA (G/A)GT ACC AGT (G/C)AT CAT GTT | ||||
| Translation elongation factor 1-alpha (tef1) | ~700 bp | Ef728M | CATCGAGAAGTTCGAGAAGG | 1. 95˚C– 3 min 2. 95˚C– 30 s 3. 58˚C– 30 s 4. 72˚C– 1 min Steps 2–4 x 35 cycles 5. 72˚C– 10 min |
[22, 23] |
| Tef1R | GCCATCCTTGGAGATACCAGC | ||||
| Small Subunit (SSU, 18S) of the rRNA | ~1200 bp | NS1 | GTA GTC ATA TGC TTG TCT C | 1. 95˚C– 5 min 2. 95˚C– 30 s 3. 52˚C– 30 s 4. 72˚C– 1 min Steps 2–4 x 35 cycles 5. 72˚C– 8 min |
[18] |
| NS4 | CTT CCG TCA ATT CCT TTA AG | ||||
| Large Subunit (LSU, 28S) of the rRNA | ~1200 bp | LROR | ACC CGC TGA ACT TAA GC | 1. 95˚C– 5 min 2. 95˚C– 30 s 3. 52˚C– 30 s 4. 72˚C– 1 min Steps 2–4 x 35 cycles 5. 72˚C– 8 min |
[24, 25] |