Table 8. Oligonucleotide primers for amplifying 38 gene fragments and temperature used for denaturing HPLC
Locus |
Primer |
Sequence, 5-3¢ |
Primer |
Sequence, 5-3¢ |
Temperature, ° C |
YPO0142 |
O3446 |
GCTGAAGGCTCAGGGTAATG |
O3447 |
CAGTACATCGGCACTGATCG |
59.8 |
YPO0163 |
O3448 |
TAGGCACCTTCTTGGTCACC |
O3449 |
ACTGACCAATTGGCCTGC |
60.6 |
YPO0185 |
O3450 |
ACAGCTATTGGCCGGGTTAC |
O3451 |
GGCGTAATGCTGTGTCACAC |
60.0 |
YPO0256 |
O4584 |
GCATCCAGTGGTGTATTTT |
O4585 |
CAGCAGCAGTGACAGTACTGA |
64.5 |
YPO0282 |
O3452 |
ACAGGCTAAAGCAGAGCACC |
O3453 |
GGAATAACCGGAGGTCCAAG |
61.8 |
YPO0285 |
O3454 |
GAATAGATCGCCACTGGCTG |
O3455 |
GTGCTCTGATATGGCCACTG |
61.5 |
YPO0326 |
O3456 |
GTGCTCCTTGTAACCCACGT |
O3457 |
GTTTGAGCCACCGCATTC |
59.6 |
YPO0462 |
O4556 |
GGAACTGCAAGGCGATAATATTC |
O4557 |
TAGTGCTGTTCAGCCCGTTTAAA |
57.9 |
YPO0643 |
O3458 |
CCAGGAAGGCAACATTGG |
O3459 |
CAGGCTCTCAGAAGTTGCAG |
60.2 |
YPO0835 |
O4582 |
GCTACGTTAGTCGGAATCTG |
O4583 |
GCATACCAACGCCCATTGC |
62.0 |
YPO0858 |
O3460 |
ATGGCTTAGTAGGATCGGGG |
O3461 |
CCCCTGGTTGGCTCATCTA |
56.2 |
YPO0898 |
O4560 |
GCGCTGGCGGAAATCTTTA |
O4561 |
GATGTCCAGTGCCAGCCACT |
62.1 |
YPO1037 |
O3594 |
AAAATAAGGTGCGTCAAAG |
O3595 |
CAGATCATCAAGGCACCGTAA |
61.5 |
YPO1224 |
O3466 |
GTTGAAACGTCTCTCTGCCC |
O3467 |
ATGCGGATCAGTGAGTCTGG |
60.6 |
YPO1246 |
O3468 |
GAACACCTTCATCGAGCCTC |
O3469 |
TCTGCGGAGCCAGTAATAGC |
60.1 |
YPO1277 |
O3470 |
GCTGTGTTAATGGGCTACCG |
O3471 |
GCCGTGATGATGTGTACCAG |
61.1 |
YPO1291 |
O4562 |
CTGGAACGACCAACGCAATA |
O4563 |
CGCAGCCGCAAGTAGTCTTT |
62.9 |
YPO1372 |
O4564 |
CCGAGTGGGTCAGCTAATGG |
O4565 |
ATTTACCACAACCAGTACGGCC |
62.1 |
YPO1402 |
O3472 |
CCTTAGGCCATCAGGTGCT |
O3473 |
GCTCAAGCCAACCGTATACC |
60.7 |
YPO1526 |
O3474 |
CTGGAAGGGGCTGATATTCC |
O3475 |
ACCGGTTAGGCGAGTTACCT |
61.5 |
YPO1574 |
O4566 |
TGAAAAAGAAAACACGATCAAACATT |
O4567 |
AGAGGTATCAATAGTAACAATCGCATAATT |
56.0 |
YPO1581 |
O4568 |
AACGATTTAAGTTCACTGGGCG |
O4569 |
TTTCAGTAGCGGGTAGCCGT |
59.2 |
YPO2180 |
O3482 |
GAACTCCTTCAAGCCAGACG |
O3483 |
CCTGGCCGTCAGAGTATTCA |
59.4 |
YPO2291 |
O3484 |
CACTTTGGGACTCCCTTGC |
O3485 |
CGAGTGGGATCTTCAGTTCC |
59.5 |
YPO2420 |
O4570 |
AAGCGATTGTATTTGCCTATCATG |
O4571 |
CTCAACCCACAAAGGGTGGT |
60.6 |
YPO2439 |
O3486 |
CCGTGGTGGAGTCAATAACC |
O3487 |
CTGTTCCGCAGGAATACGAG |
62.3 |
YPO2612 |
O3488 |
CAGGTGCTGGCTGACTGTAC |
O3489 |
CCTTTCTCAAGGGCTTCTGC |
58.4 |
YPO2676 |
O4572 |
CGGTACCACGTCTGGCTGA |
O4573 |
TTTTCATCATTTCTACTGCGCC |
58.1 |
YPO2875 |
O4574 |
TGGCAGAGGAAGACCATCAAC |
O4575 |
ACTGAATGCGGATTGGCG |
60.0 |
YPO3120 |
O4576 |
AAGCCCGGCAGATATCCAT |
O4577 |
GCTGTTGCATCCCCTTCAAC |
60.2 |
YPO3268 |
O4578 |
CTAGCCAAGCGAGTCGGTG |
O4579 |
CATACTTCGCTTCGCCGG |
63.0 |
YPO3339 |
O3490 |
GATCACTACAGCTTGCCCGT |
O3491 |
CTGCCCTTGCTCCGTATCT |
56.4 |
YPO3352 |
O3492 |
AGGGTATCCCCGAGACCAT |
O3493 |
GGAAGCCGTACAGATCGTGT |
60.3 |
YPO3841 |
O3498 |
GGCAACTTGTTGGTACTGGC |
O3499 |
CCGGATTACCGTAGCTCTCA |
62.4 |
YPO3897 |
O3500 |
GTACCGTTCCCATACCACCA |
O3501 |
AGAGTAGAGCATGCCACC |
61.7 |
YPO3943 |
O3502 |
CCTTCAATGAAGCGGTCG |
O3503 |
AACCCAGCTTACAGGTGGGT |
62.1 |
YPO4007 |
O3504 |
GTCTGCGAGATAAGCCAAGC |
O3505 |
AGAAGCAGCCTGCCTGTAAG |
58.7 |
YPO4066 |
O4580 |
CACCCGGCATGAATATGAAGAT |
O4581 |
TGGCGTTGCATAGCAATCAG |
61.4 |
For YPO0256 and YPO1574 gradients were performed by using a time shift of 1.3 and 0.5 min, respectively.