Table 1 Primers and Probes and observed melting temperatures employed for FCN2 SNP detection
Position | Primer/probe (Conc.) | Oligonucleotide sequence (5′- 3′) | Genotypes (Tm) |
|---|---|---|---|
−986 G > A (rs3124952) | Forward (0.5 μM) | 5′-GGGTCACAGTTTAAAATCCTTCTACT-3′ | |
Reverse (0.05 μM) | 5′-CGTATACCTAAAGCCCCCAGA-3′ | GG (73°C) | |
Sensor (0.25 μM) | CY5-GCCACCTGC[C]GCCATCG-PH | GA (73°C/64°C) | |
Anchor (0.25 μM) | 5′CCTCCCACTACCACCACCGCACCC-FL | AA (64°C) | |
−602 G > A (rs3124953) | Forward (0.05 μM) | 5′-CAAGGTCTCCCCTTCAGATG-3′ | |
Reverse (0.5 μM) | 5′-CATGAGCAGACTTGGGACT-3′ | GG (60°C) | |
Sensor (0.15 μM) | 5′-CCTCCTGTTC[A]TGTGCCCC-FL 3′ | GA (60°C/67°C) | |
Anchor (0.15 μM) | CY5-GTGCTCTACATACTGCCCCAGGAAACAG-PH | AA (67°C) | |
−4 A > G (rs17514136) | Forward (0.125 μM) | 5′-GGAAGCGGCTGTCACTC-3′ | |
Reverse (0.5 μM) | 5′-CCCTTACCTGGACAGGTGT-3′ | AA (64°C) | |
Sensor (0.15 μM) | 5′AGCAAAGACCAGA[A]GAGATGGA-FL | AG (64°C/58°C) | |
Anchor (0.15 μM) | CY5-CTGGACAGAGCTGTGGGGGTC-PH | GG (58°C) | |
+6424 G > T (rs7851696) | Forward (0.5 μM) | 5′-TGCCTGTAACGATGCTCA-3′ | |
Reverse (0.05 μM) | 5′-TGTATCCTTTCCCCGACTT-3′ | GG (65°C) | |
Sensor (0.15 μM) | 5′-GAAACATCACAG[C]ACAATTTCC-FL | GT (65°C/55°C) | |
Anchor (0.15 μM) | CY5-GTGTTAAGATCATTGTCCTGGTCTTTGGT-PH | TT (55°C) |