Table 3.
Primer Sequences and PCR Amplification Conditions for Validated Genetic Variants before Performing Verification by Sanger Sequencing
| Gene | rs number | HGVS nomenclature | Sequence accession number | Forward primer sequence | Reverse primer sequence | PCR annealing temperature, degrees Celsius | PCR product/amplicon length, bp |
|---|---|---|---|---|---|---|---|
| ABCB1 | rs1045642 | c.3435T>C or p.Ile1145= | NM_000927.4 or NP_000918.2 | 5′-ACTCTTGTTTTCAGCTGCTTG-3′41 | 5′-AGAGACTTACATTAGGCAGTGACTC-3′41 | 63 | 231 |
| rs1128503 | c.1236T>C or p.Gly412= | 5′-TGTGTCTGTGAATTGCCTTGAAG-3′42 | 5′-CCTCTGCATCAGCTGGACTGT-3′42 | 63 | 149 | ||
| rs2032582 | c.2677T>G/A or p.Ser893Ala/Thr | 5′-ATGGTTGGCAACTAACACTGTTA-3′42 | 5′-AGCAGTAGGGAGTAACAAAATAACA-3′42 | 63 | 208 | ||
| CBR3 | rs1056892 | c.730G>A or p.Val244Met | NM_001236.3 or NP_001227.1 | 5′-CCAGGACCAGTGAAGACAGA-3′ | 5′-CCGAAGCAGACGTTTACCAG-3′ | 63 | 166 |
| COMT | rs4646316 | c.615+310C>T | NM_000754.3 | 5′-ACACGCTTCTCTTGGAGGTG-3′ | 5′-CTGTCTAGCCTCACTCGGG-3′ | 63 | 519 |
| rs9332377 | c.616-367C>A/T | 5′-GCTTGTTGATGGGAGGTCTG-3′ | 5′-TCCCTTAGAACAGCATGTGG-3′ | 61 | 217 | ||
| C8ORF34 | rs1517114 | c.736+8162C>G/T/A | NM_052958.2 | 5′-CTGTGCTTTCTCGTCTTCAG-3′ | 5′-CAGCCTGGAACCTACCCTTG-3′ | 58 | 238 |
| FCGR2A | rs1801274 | c.500A>G or p.His167Arg | NM_001136219.1 or NP_001129691.1 | 5′-CAAGCCTCTGGTCAAGGTCA-3′ | 5′-AAGGATTCCCCTTAGCCCCT-3′ | 58 | 663 |
| FCGR3A | rs396991 | c.841T>C/G or p.Phe281Leu/Val | NM_000569.7: or NP_000560.6 | 5′-CACATATTTACAGAATGGCAAAGG-3′ | 5′-GATTCTGGAGGCTGGTGCTACA-3′ | 58 | 969 |
| HAS3 | rs2232228 | c.279A>G or p.Ala93= | NM_001199280.1 or NP_001186209.1 | 5′-GTGACGGGCTACCAGTTCAT-3′ | 5′-CACAACCCAAGGGACCTAGA-3′ | 58 | 654 |
| NT5C2 | rs11598702 | c.175+1178A>G/C | NM_012229.4 | 5′-GACGGGTTTATAGGTGCAGC-3′ | 5′-TCAATGACTTCTTGCCCAGT-3′ | 58 | 222 |
| NUDT15 | rs116855232 (*3) | c.415C>T or p.Arg139Cys | NM_018283.3 or NP_060753.1 | 5′-GCCTTTGTAAACTGGGCTTC-3′ | 5′-CAAATCTTCTCGGCCACCTA-3′ | 58 | 411 |
| rs186364861 (*5) | c.52G>A or p.Val18Ile | 5′-CATTCCCCAACCTGATAGCC-3′ | 5′-CAACCGAGCCTTTCCTCTTC-3′ | 58 | 296 | ||
| SBF2 | rs7102464 | c.2035G>A or p.Glu679Lys | NM_030962.3 or NP_112224.1 | 5′-ACAGAAACTTGCCCCTGGAG-3′ | 5′-ACCCAAATACACTGGCAGGA-3′ | 63 | 289 |
| rs146987383 | c.2050C>G or p.Leu684Val | 5′-ACAGAAACTTGCCCCTGGAG-3′ | 5′-ACCCAAATACACTGGCAGGA-3′ | 63 | 289 | ||
| rs141368249 | c.2081C>T or p.Ala694Val | 5′-ACAGAAACTTGCCCCTGGAG-3′ | 5′-ACCCAAATACACTGGCAGGA-3′ | 63 | 289 | ||
| rs117957652 | c.3292C>G/T or p.Leu1098Val/= | 5′-CCTGTCTTGGTGTAAGAGTCTTCT-3′ | 5′-ACCTCTTTTTGGAGCCCACT-3′ | 63 | 843 | ||
| rs149501654 | c.4111G>C or p.Val1371Leu | 5′-TCTTCATCCGCAGAACTTCA-3′ | 5′-AGTGTGCCTTTGGTGGGTAG-3′ | 63 | 649 | ||
| SEMA3C | rs7779029 | c.103+13883A>G | NM_006379.3 | 5′-GGCTTAGGTCTCTGCCCTTT-3′ | 5′-GTTCCCATTTCCAGGCTCCA-3′ | 58 | 200 |
| rs11979430 | c.103+36739G>A | 5′-GGAAAGGGCAGACTGTGGTA-3′ | 5′-ACCAAACCTCTTCAGGGTGA-3′ | 58 | 383 | ||
| SLC16A5 | rs4788863 | c.121T>C or p.Leu81= | NM_004695.3 or NP_004686.1 | 5′-AGGTCCCCCTGTTGACTTCT-3′ | 5′-TGAAATCTGGTGAAACCTTAGGA-3′ | 58 | 725 |
| SLC28A3 | rs885004 | c.862-360C>T | NM_022127.2 | 5′-TGTGTCTGCCATCCAGTAGG-3′ | 5′-CCTGGTGCTAAAAAGACATGG-3′ | 58 | 161 |
| rs7853758 | c.1381C>T or p.Leu461= | NM_022127.2 or NP_071410.1 | 5′-CCCCTGACAACTCCTTGGTA-3′ | 5′-CAGGGGCGTGATGTGATTAT-3′ | 58 | 239 | |
| SOD2 | rs4880 | c.47T>C or p.Val16Ala | NM_000636.3 or NP_000627.2 | 5′-CTGTGCTTTCTCGTCTTCAG-3′ | 5′-CAGCCTGGAACCTACCCTTG-3′ | 58 | 238 |
| TLR4 | rs4986790 | c.776A>G or p.Asp299Gly | NM_003266.3 or NP_612564.1 | 5′-AGTCCATCGTTTGGTTCTGG-3′ | 5′-TGCCATTGAAAGCAACTCTG-3′ | 58 | 635 |
| TPMT | rs12201199 | c.419+94T>A | NM_000367.3 | 5′-GTTCTTCGGGGAACATTTCA-3′ | 5′-AAGTGATTGAGCCACAAGCC-3′ | 58 | 975 |
Accession numbers are available at https://www.ncbi.nlm.nih.gov/snp.
HGVS, Human Genome Variation Society.