Primers used in the ARMS reactions to genotype codons 198, 219, 234, 259, and 376 and the resulting amino acid in transcobalamin
| Codons . | Forward primer containing the point mutation at the 3′ end and the resulting amino acid . | Reverse primer . | Molecular weight . |
|---|---|---|---|
| 198 | PIS:5′ acttcaag 581 AC ACA GCA GCA AT 593 3′ → Met | ASP: 5′805 TGA GAC ATG CTG TTC CCA GTT 785 3′ | 358 base pairs (bp) |
| TACH: 5′ acttcaag581 AC ACA GCA GCA AC 593 3′→ Thr | |||
| 219 | RAI: 5′637 CCT GGT CGG AGA CAA CAG A 655 3′→ Ile | ASP: 5′ 805 TGA GAC ATG CTG TTC CCA GTT 785 3′ | 294 bp |
| SIN: 5′ 637 CCT GGT CGG AGA CAA CAG C 655 3′ → Leu | |||
| 234 | CIT: 5′682 GAG GAG ATC TTG AAG GCA CA 701 3′→ Gln | ASP: 5′ 805 TGA GAC ATG CTG TTC CCA GTT 785 3′ | 249 bp |
| RON: 5′ 682 GAG GAG ATC TTG AAG GCA CG 701 3′ →Arg | |||
| 259 | CAS: 5′757 CTC ATG ACT TCC CCC ATG CC 776 3′→ Pro | ERGE: 5′ 933 TGC CAG ACA GTC TGG GAA GA 914 3′ | 176 bp |
| SIS: 5′ 757 CTC ATG ACT TCC CCC ATG CG 776 3′ →Arg | |||
| 376 | CHOC: 5′1108 TAT GAA ACA CAG GCC TCC TC 1127 3′→ Ser | AS4: 5′ 1230 AGC AAT ACC TTG CAA CAG TGG 1210 3′ | 122 bp |
| OLA: 5′ 1108 TAT GAA ACA CAG GCC TCC TT 1127 3′ → Leu |
| Codons . | Forward primer containing the point mutation at the 3′ end and the resulting amino acid . | Reverse primer . | Molecular weight . |
|---|---|---|---|
| 198 | PIS:5′ acttcaag 581 AC ACA GCA GCA AT 593 3′ → Met | ASP: 5′805 TGA GAC ATG CTG TTC CCA GTT 785 3′ | 358 base pairs (bp) |
| TACH: 5′ acttcaag581 AC ACA GCA GCA AC 593 3′→ Thr | |||
| 219 | RAI: 5′637 CCT GGT CGG AGA CAA CAG A 655 3′→ Ile | ASP: 5′ 805 TGA GAC ATG CTG TTC CCA GTT 785 3′ | 294 bp |
| SIN: 5′ 637 CCT GGT CGG AGA CAA CAG C 655 3′ → Leu | |||
| 234 | CIT: 5′682 GAG GAG ATC TTG AAG GCA CA 701 3′→ Gln | ASP: 5′ 805 TGA GAC ATG CTG TTC CCA GTT 785 3′ | 249 bp |
| RON: 5′ 682 GAG GAG ATC TTG AAG GCA CG 701 3′ →Arg | |||
| 259 | CAS: 5′757 CTC ATG ACT TCC CCC ATG CC 776 3′→ Pro | ERGE: 5′ 933 TGC CAG ACA GTC TGG GAA GA 914 3′ | 176 bp |
| SIS: 5′ 757 CTC ATG ACT TCC CCC ATG CG 776 3′ →Arg | |||
| 376 | CHOC: 5′1108 TAT GAA ACA CAG GCC TCC TC 1127 3′→ Ser | AS4: 5′ 1230 AGC AAT ACC TTG CAA CAG TGG 1210 3′ | 122 bp |
| OLA: 5′ 1108 TAT GAA ACA CAG GCC TCC TT 1127 3′ → Leu |
The point mutation of interest occurs at the 3′ terminus last base of the forward primers (bold). The additional mismatch introduced in some primers is underlined.