Table 1.
List of primers used for the three-step PCR methodology.
| Primer | Sequence | PCR Step |
|---|---|---|
| ITS4 | 5′TCCTCCGCTTATTGATATGC 3′ | 1 |
| ITS1F | 5′CTTGGTCATTTAGAGGAAGTAA 3′ | |
| LROR | 5′ACCCGCTGAACTTAAGC 3′ | |
| LR3 | 5′CCGTGTTTCAAGACGGG 3′ | |
| 806R | 5′GGACTACHVGGGTWTCTAAT 3′ | |
| 341F | 5′CCT ACG GGN GGC WGC AG 3′ | |
| ITS4 f1 | 5′GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNNNNAATCCTCCGCTTATTGATATGC 3′ | 2 |
| ITS4 f2 | 5′GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNTNNNAATCCTCCGCTTATTGATATGC 3′ | |
| ITS4 f3 | 5′GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNCTNNNAATCCTCCGCTTATTGATATGC 3′ | |
| ITS4 f4 | 5′GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNACTNNNAATCCTCCGCTTATTGATATGC 3′ | |
| ITS4 f5 | 5′GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNGACTNNNAATCCTCCGCTTATTGATATGC 3′ | |
| ITS4 f6 | 5′GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNTGACTNNNAATCCTCCGCTTATTGATATGC 3′ | |
| ITS1F f1 | 5′GCCTCCCTCGCGCCATCAGAGATGTGTATAAGAGACAGNNNNNNNNTTCTTGGTCATTTAGAGGAAGTAA 3′ | |
| ITS1F f2 | 5′GCCTCCCTCGCGCCATCAGAGATGTGTATAAGAGACAGNNNNTNNNNTTCTTGGTCATTTAGAGGAAGTAA 3′ | |
| ITS1F f3 | 5′GCCTCCCTCGCGCCATCAGAGATGTGTATAAGAGACAGNNNNCTNNNNTTCTTGGTCATTTAGAGGAAGTAA 3′ | |
| ITS1F f4 | 5′GCCTCCCTCGCGCCATCAGAGATGTGTATAAGAGACAGNNNNACTNNNNTTCTTGGTCATTTAGAGGAAGTAA 3′ | |
| ITS1F f5 | 5′GCCTCCCTCGCGCCATCAGAGATGTGTATAAGAGACAGNNNNGACTNNNNTTCTTGGTCATTTAGAGGAAGTAA 3′ | |
| ITS1F f6 | 5′GCCTCCCTCGCGCCATCAGAGATGTGTATAAGAGACAGNNNNTGACTNNNNTTCTTGGTCATTTAGAGGAAGTAA 3′ | |
| LROR f1 | 5′GCCTCCCTCGCGCCATCAGAGATGTGTATAAGAGACAGNNNNNNNNGAACCCGCTGAACTTAAGC 3′ | |
| LROR f2 | 5′GCCTCCCTCGCGCCATCAGAGATGTGTATAAGAGACAGNNNNTNNNNGAACCCGCTGAACTTAAGC 3′ | |
| LROR f3 | 5′GCCTCCCTCGCGCCATCAGAGATGTGTATAAGAGACAGNNNNCTNNNNGAACCCGCTGAACTTAAGC 3′ | |
| LROR f4 | 5′GCCTCCCTCGCGCCATCAGAGATGTGTATAAGAGACAGNNNNACTNNNNGAACCCGCTGAACTTAAGC 3′ | |
| LROR f5 | 5′GCCTCCCTCGCGCCATCAGAGATGTGTATAAGAGACAGNNNNGACTNNNNGAACCCGCTGAACTTAAGC 3′ | |
| LROR f6 | 5′GCCTCCCTCGCGCCATCAGAGATGTGTATAAGAGACAGNNNNTGACTNNNNGAACCCGCTGAACTTAAGC 3′ | |
| LR3 f1 | 5′GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNNNNCACCGTGTTTCAAGACGGG 3′ | |
| LR3 f2 | 5′GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNTNNNCACCGTGTTTCAAGACGGG 3′ | |
| LR3 f3 | 5′GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNCTNNNCACCGTGTTTCAAGACGGG 3′ | |
| LR3 f4 | 5′GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNACTNNNCACCGTGTTTCAAGACGGG 3′ | |
| LR3 f5 | 5′GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNGACTNNNCACCGTGTTTCAAGACGGG 3′ | |
| LR3 f6 | 5′GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNTGACTNNNCACCGTGTTTCAAGACGGG 3′ | |
| 806R f1 | 5′GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNNNNACGGACTACHVGGGTWTCTAAT 3′ | |
| 806R f2 | 5′GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNTNNNACGGACTACHVGGGTWTCTAAT 3′ | |
| 806R f3 | 5′GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNCTNNNACGGACTACHVGGGTWTCTAAT 3′ | |
| 806R f4 | 5′GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNACTNNNACGGACTACHVGGGTWTCTAAT 3′ | |
| 806R f5 | 5′GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNGACTNNNACGGACTACHVGGGTWTCTAAT 3′ | |
| 806R f6 | 5′GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNTGACTNNNACGGACTACHVGGGTWTCTAAT 3′ | |
| 341F_f1 | 5′GCC TCC CTC GCG CCA TCA GAG ATG TGT ATA AGA GAC AGN NNN NNN NAG CCT ACG GGN GGC WGC AG 3′ | |
| 341F_f2 | 5′GCC TCC CTC GCG CCA TCA GAG ATG TGT ATA AGA GAC AGN NNN TNN NNA GCC TAC GGG NGG CWG CAG 3′ | |
| 341F_f3 | 5′ GCC TCC CTC GCG CCA TCA GAG ATG TGT ATA AGA GAC AGN NNN CTN NNN AGC CTA CGG GNG GCW GCA G 3′ | |
| 341F_f4 | 5′GCC TCC CTC GCG CCA TCA GAG ATG TGT ATA AGA GAC AGN NNN ACT NNN NAG CCT ACG GGN GGC WGC AG 3′ | |
| 341F_f5 | 5′GCC TCC CTC GCG CCA TCA GAG ATG TGT ATA AGA GAC AGN NNN GAC TNN NNA GCC TAC GGG NGG CWG CAG 3′ | |
| 341F_f6 | 5′GCC TCC CTC GCG CCA TCA GAG ATG TGT ATA AGA GAC AGN NNN TGA CTN NNN AGC CTA CGG GNG GCW GCA G 3′ | |
| PCR_F | 5′AATGATACGGCGACCACCGAGATCTACACGCCTCCCTCGCGCCATCAGAGATGTG 3′ | 3 |
| PCR_R_bc | 5′CAAGCAGAAGACGGCATACGAGAT XXXXXXXXXGTGACTGGAGTTCAGACGTGTGCTC 3′ 1 |
1 = XXX on the primer sequence represents a specific barcode per sample.