TABLE 3.
Oligonucleotides used in this study
| Primer | Sequence (5′ → 3′) |
|---|---|
| Mutagenesis | |
| HIS4-g3c.F | TTTTTTCTGAATAATCGTTTTGCCGATTCTACCG |
| HIS4-g3c.R | CGGTAGAATCGGCAAAACGATTATTCAGAAAAAA |
| HIS4.g3c-adaptamerA.F | AATTCCAGCTGACCACCATGTATGACTATGAACAGTAG |
| HIS4.g3c-adaptamerB.R | GATCCCCGGGAATTGCCATGACCAACCCTAGACAACGCTC |
| HIS4.c1605g.F | CCAGCTAGAACAATCTTGGAAGCCCCAACTTTTTCTGCGAC |
| HIS4.c1605g.R | GTCGCAGAAAAAGTTGGGGCTTCCAAGATTGTTCTAGCTGG |
| HIS4-1605-MX4.F | TGTCTTGTGTTCCAGATTCCCTCGTCCTATTGAAAAAGTTGGCGTACGCTGCAGGTCGAC |
| HIS4-1605-MX4.R | CCAAAACTTCACTTGGGCCAGCTGGCATATCAATGGAACATAGAGCATCGATGAATTCGAGCTCG |
| BIK1.g939a.F | CGAACTCCTGAAAAAACAGCTAGAACAATTACGCAAC |
| BIK1.g939a.R | GTTGCGTAATTGTTCTAGCTGTTTTTTCAGGAGTTCG |
| BIK1.g939a-adaptamerA.F | GAACTGACTCTAATAGTGACTCCGGTAAATTAGTTAATTAATTGCGATGATGTAGTTTCTGGTT |
| BIK1.g939a-adaptamerB.R | ATGTATGTACAACACACATCGGAGGTGAATATAACGTTCCGTGATTCTGGGTAGAAGATCG |
| Cloning | |
| HIS4-1000 upstream.F | GAATTACGAGAAGCCCAATTGACCATCGAA |
| HIS4-ORF.F | ATGGTTTTGCCGATTCTACCGTTAATTGAT |
| HIS4-ORF.R | CTACTGGAAATCCTTTGGGAACAACCCAAGC |
| BIK1-500 upstream.F | AGTTGCGTTTGGGAAGAA |
| BIK1-223 downstream.R | ACAGCGCAGTTGTGCTATGATAT |
| Insertion | |
| FUS1-HPH.F | TTGTCATGCACATCATCATACTAAACTTACACGAATAGGAATCGATGAATTCGAGCTCG |
| FUS1-HPH.R | TTGTCATGCACATCATCATACTAAACTTACACGAATAGGAATCGATGAATTCGAGCTCG |
| FUS1-A1 | CATTGCCGCTTACTCCAAAC |
| FUS1-A4 | CATTGCCGCTTACTCCAAAC |
| RRP7-NAT.F | TTTAAAGGCATCAATAATTTTTTTCTTTCATATATATTCTGCGTACGCTGCAGGTCGAC |
| RRP7-NAT.R | TTGTCATGCACATCATCATACTAAACTTACACGAATAGGAATCGATGAATTCGAGCTCG |
| RRP7-A1 | GTGGATGAGGATGGATTCAC |
| RRP7-A4 | CATAACCGACAATCACCGTC |
| Allele detection | |
| HIS4.1200-1220.fwd | TCTGTTTTTTTGGAGTACAC |
| HIS4.1700-1681.rev | GGACCCAAGATCTTATCCAC |
| BIK1.751-771.fwd | AAGCAACAATTGGAGCTCGAACGC |
| BIK1.1323-1300.R | CTAGAAGAACTGCTGGTTGTCAGG |
Disruption oligonucleotide primers for MLH1, MSH2, and COM1 were designed using the immediate 40–45 bp upstream and downstream and linking these to the KANMX4 primer sequence. Verification primers were designed upstream and downstream as described previously (Wach et al. 1994). Sequences are available upon request.