Primers and conditions for detection of the common glomulin mutations by allele specific polymerase chain reaction or restriction enzyme digestion
| 108C→A | 157delAAGAA | 554delA+556delCCT | 1179delCAA | |
|---|---|---|---|---|
| *Only 50% of this primer should be used in the allele specific polymerase chain reaction. | ||||
| | = site of deletion; underlined nucleotide = substitution. | ||||
| bp, base pair; temp, temperature. | ||||
| Forward primer | GATACGTGTGTTATTTACGTTAC | GATACGTGTGTTATTTACGTTAC | TTGATGAGCGACAACTTGATC | GTGATGAAGTCTGGGTAAGC |
| Reverse primer | ATGTGATTATTCTCTTCCCAAG | ATGTGATTATTCTCTTCCCAAG* | TAAGTCCACTGTGAGATGTTC | AACAATTACATGGCATTAACATG |
| Wt product size | 324 bp | 324 bp | 304 bp | 207 bp |
| Mutation specific ( = 3rd) primer | F: AGTTAGCTGGGCAAAGATGA | R: GTACGCACCTTA|TTCATTTTG | – | R: TTGCCTTGTGAATCCAAC|TTAA |
| Restriction enzyme | – | – | Spe I | – |
| Mutant product(s) | 130 bp | 219 bp | 111 and 193 bp | 117 bp |
| Annealing temp | 55°C | 55°C | 62°C | 58°C |