Allele/SNP | Primer Name | Primer Sequence | Annealing Temperature | Restriction Enzyme | Digestion Products |
---|---|---|---|---|---|
°C | |||||
Cyp3A5*2 | 5Ex11f | CTTCACGAATACTATGATCATTTACC | 60 | Tsp5091 (NEB) | Wt: 310,90,22 |
5Ex11r | GGCAGAATATGCTTGAACCAGC | Mt:187,123,90,22 | |||
Cyp3A5*3 | 3A5A3f | CATGACTTAGTAGACAGATGA | 56 | Ssp I (NEB) | Wt: 148,125,20 |
3A5A3r | GGTCCAAACAGGGAAGAAATA | Mt: 168,125 | |||
Cyp3A5*5 | 3A5A5f | CCATGAAGATCACCACAACT | 56 | Nla III (NEB) | Wt: 218 |
3A5A5r | CCTGTCCCCAGATTCATTC | Mt: 172,46 | |||
Cyp3A5*6 | 5Ex7f | GCATGTATAGTGGAAGGACG | 60 | Dde I (NEB) | Wt: 103,60,87,25 |
5Ex7r | GAATTATATGTCAAGAAAGCAG | Mt: 128,87,60 | |||
SNP 31611 | C3A512f | ACCCTAAGTGGAGGAATGAGT | 60 | Sca 1 (NEB) | Wt: 209 |
(3′-Untranslated region) | TATTCTAAGT | Mt: 181,29 | |||
| 5Ex13r | TCCTTTATACTACATAATGCACAAC |
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NEB, New England Biolabs (Beverly, MA); Wt, wild-type; Mt, mutant.