Table 1 Oligonucleotide primers for amplification of X. laevis ADHs
Name | Nucleotide sequence1 | Amino acid residues2 |
|---|---|---|
Degenerate primers (based on Rana perezi ADH8) | ||
Degenerate forward outer | 5′-ATGTGCACTGCGGGIAARGWIATHA-3′ | 1-8 |
Degenerate reverse outer | 5′-TAGTCTTTTGGRTTIADRCAYTC-3′ | 237-244 |
Degenerate forward inner | 5′-ATTACATGTAAGGCIGCIGTIGC-3′ | 7-14 |
Degenerate reverse inner | 5′-GGCTTTTGGRAAYTTRTCYTTRT-3′ | 223-230 |
Specific forward primers (used in combination with 3′-end adaptors) | ||
ADH1B outer | 5′-GTATAGTGGAAAGTGTGGGAGAG-3′ | 72-79 |
ADH1B inner | 5′-CATACATTGGACTCTTGTTGGAC-3′ | 119-126 |
ADH3 outer | 5′-CTGAATACACTGTTGTAGC-A-3′ | 148-154 |
ADH3 inner | 5′-CAACTGGTTATGGAGCTGTG-3′ | 178-184 |
ADH8B outer | 5′-AGCACTTTTACAGAATACAG-3′ | 144-150 |
ADH8B inner | 5′-AGATTCCTCCAGGATCTACG-3′ | 186-192 |
ADH9 outer | 5′-GCTAAAGTACAGCAAGGTAG-3′ | 189-194 |
ADH9 inner | 5′-GAATCATTGGAGTAGACATT-3′ | 219-225 |
Specific reverse primers for ADH8B (used in combination with the above specific forward primers) | ||
ADH8B reverse outer | 5′-TCAACAGGATGTCAGGCTGCAAATG-3′ | Nucleotides 61–37 of non-coding 3′-end |
ADH8B reverse inner | 5′-AATGACCGTAGTGGACTTCACACGA-3′ | Nucleotides 40–16 of non-coding 3′-end |