Polymerase chain reaction (PCR) primer sequences, PCR amplification conditions, and lengths for each of the candidate SNP amplimers
| Locus | Amplimer length | Primer sequences | Cycling conditions |
|---|---|---|---|
| Pyrosequencing primer sequences are also presented. | |||
| TNF-α −308 | 107 bp | PCR (For): AGGCAATAGGTTTTGAGGGCCAT | One cycle: 95°C - 4 min; 35 cycles: 95°C - 3 s, |
| PCR (Rev): TCCTCCCTGCTCCGATTCCG | 60°C - 30 s, 74°C - 30 s; one cycle: 74°C - 6 min | ||
| Pyrosequencing (Rev): GGCTGAACCCCGTCC | |||
| IL-6 −174 | 111 bp | PCR (For): CGCTAGCCTCAATGAC | One cycle: 95°C - 4 min; 35 cycles: 95°C - 45 s, |
| PCR (Rev): CGGGTGGGGCTGATTGGAA | 63°C - 30 s, 74°C - 45 s; one cycle: 74°C - 6 min | ||
| Pyrosequencing (For): CCCTAGTTGTGTCTTGC | |||
| IL-1β −31 | 222 bp | PCR (For): CCTCCTACTTCTGCTTTTGAAGGC | One cycle: 95°C - 4 min; 35 cycles: 95°C - 30 s, |
| PCR (Rev): AAGAGAGAGAGAGAGAGAATATGC | 55°C - 30 s, 74°C - 30 s; one cycle: 74°C - 6 min | ||
| Pyrosequencing (Rev): TCCCTCGCTGTTTTTA | |||
| CD14 −159 | 193 bp | PCR (For): ATCATCCTTTTCCCACACC | One cycle: 95°C - 4 min; 35 cycles: 95°C, 30 s, |
| PCR (Rev): GCCCCCTTCCTTTCCTGGA | 64°C - 30 s, 74°C - 30 s; one cycle: 74°C - 6 min | ||
| Pyrosequencing (Rev): TTCAGGGAGGGGG | |||
| TLR4 +896 | 193 bp | PCR (For): AGTCCATCGTTTGGTTCTGG | One cycle: 95°C - 4 min; 35 cycles: 95°C - 60 s, |
| PCR (Rev): CACACTCACCAGGGAAAATG | 55°C - 60 s, 74°C - 60 s; one cycle: 74°C - 6 min | ||
| Pyrosequencing (Rev): ACAATTAAATAAGTCAATAA | |||