Primer sequences used for the pyrosequencing-based genotyping of functional BCRP variants
SNPa | Variant | Primer Sequence (5′→3′) | Size | PCR Condition |
|---|---|---|---|---|
| base pair | Tm; °C | |||
| 34G>A | V12M | 5′-Biotin-CTCTCCAGATGTCTTCCAGTAATG-3′ | 278 | 54.0 |
| 5′-GCCAAAACCTGTGAGGTTCA-3′ | ||||
| For sequencing: 5′-AGTGTTCCTTTGTGGTTAC-3′ | ||||
| 8191C>T | Q126Stop | 5′-Biotin-ACTATCAGCCAAAGCACTTACCC-3′ | 174 | 54.5 |
| 5′-GTCTTAGCTGCAAGGAAAGATCCA-3′ | ||||
| For sequencing: 5′-AATGTAATTCAGGTTACGTG-3′ | ||||
| 8825C>A | Q141K | 5′-Biotin-GTTGCAAGCCGAAGAGCTG-3′ | 69 | 54.0 |
| 5′-TGATGTTGTGATGGGCACTC-3′ | ||||
| For sequencing: 5′-GACGGTGAGAGAAAACTT-3′ | ||||
| 21850C>T | P269S | 5′-Biotin-TAGCACCAAATGGAACAAACAC-3′ | 236 | 54.0 |
| 5′- TGTTTGATAGCCTCACCTTATTGG-3′ | ||||
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| For sequencing: 5′-GAAGACTTATGTTCCACG-3′ |
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↵ a Position is indicated with respect to the start codon (ATG) of the BCRP gene; the A in the ATG triplet is designated as +1, and the next base toward the 5′-end is designated as -1.