TABLE 1.
Primer sequence and PCR-RFLP assay conditions for genotyping SNPs of Gal2-HpyCH4IV, Gal3-Aval, Gal4-AluI, Gal5-HinfI, and Gal7-MlyI
| Gene (GenBank accession no.) | Primer sequences (forward/reverse) | PCR product size (bp) | PCR digested sizes (bp) | Annealing Temp (°C)/time (s)a | Position (SNP)b | Restriction enzyme |
|---|---|---|---|---|---|---|
| Gal2 (AY621317) | 5′-GGCACAAAGGGTAAAGTATGG-3′ | 583 | 388 + 195 | 55.1/30 | 196 (T/C) | HpyCH4IV |
| 5′-GAGGGGTCTTCTTGCTGCTGA-3′ | ||||||
| Gal3 (AY621318) | 5′-GCACCACAAGAAGCCCAGGAA-3′ | 664 | 443 + 221 | 57.3/30 | 222 (T/C) | AvaI |
| 5′-AACTCCAGCCCTTACCACTCA-3′ | ||||||
| Gal4 (AY621319) | 5′-TGGGGATCTTAGAGGTCTTTT-3′ | 600 | 416 + 184 | 51.0/30 | 188 (A/G) | AluI |
| 5′-TTTTCCACAGATATTGCTTTT-3′ | ||||||
| Gal5 (AY621320) | 5′-CTCCCAGCAAGAAAGGAACCTG-3′ | 623 | 402 + 133 + 79 | 59.0/30 | 80 (C/A) | HinfI |
| 5′-CACAGTCCTGGGGTAATCCTCG-3′ | ||||||
| Gal7 (AY621322) | 5′-CTCAGTCGGGAGATAACCATTC-3′ | 785 | 605 + 180 | 56.1/30 | 606 (G/A) | MlyI |
| 5′-GGAGTGCCAGAGAAGCCATTTG-3′ |
a
PCR annealing temperature and time for primers.
b
SNP location within the PCR fragment.