Table 4 PCR conditions used in genotyping the inflammatory gene variants
Gene | rs No | Labeled primers | Sequence(5'→3') | Amplicon (F1/R)/(F2/R) (bp) | Annealing temp.(°C) | Mg(mM) | |
|---|---|---|---|---|---|---|---|
Tumor necrosis factor-α (TNF-α) | rs1800630 | F1(FITC) | ATG GCC CTG TCT TCG TTA AxT G | 75/77 | 62.5 | 3 | |
F2(Texas Red) | GGC CCT GTC TTC GTT AAx GG | ||||||
R(Biotin) | ACA GCA ATG GGT AGG AGA ATG TC | ||||||
Interleukin-1A (IL1A) | rs1800587 | F1(FITC) | AATAATAGTAACCAGGCAACxTC | 91/92 | 60 | 2.5 | |
F2(Texas Red) | TAATAATAGTAACCAGGCAACxCC | ||||||
R(Biotin) | AGTGGCTAAGTTTGGGAATGG | ||||||
Interleukin-1B (IL1B) | rs16944 | F1(FITC) | GGTGCTGTTCTCTGCCTxGG | 50 | 60 | 1.5 | |
F2(Texas Red) | GGTGCTGTTCTCTGCCTxAG | ||||||
R(Biotin) | TCAGAGGCTCCTGCAATTGAC | ||||||
Interleukin 4 receptor (IL-4R) | rs1801275 | F1(FITC) | GCC CCC ACC AGT GGC TAT xGG | 49/50 | 67.5 | 2.8 | |
F2(Texas Red) | GGC CCC CAC CAG TGG CTA TxA G | ||||||
R(Biotin) | CAC CCT GCT CCA CCG CAT GTA | ||||||
Interleukin-6 (IL-6) | rs1800796 | F1(FITC) | GGC AGT TCT ACA ACA GCx CC | 43/42 | 60 | 1.5 | |
F2(Texas Red) | GCA GTT CTA CAA CAG CxG C | ||||||
R(Biotin) | CTG TGT TCT GGC TCT CCC TG | ||||||
Interleukin-10 (IL-10) | rs1800872 | F1(FITC) | CAG AGA CTG GCT TCC TAC AxG A | 51/53 | 62.5 | 2.5 | |
F2(Texas Red) | CCA GAG ACT GGC TTC CTA CAx TA | ||||||
R(Biotin) | GCC TGG AAC ACA TCC TGT GA | ||||||
Interleukin 1 receptor-associated kinase-1 (IRAK-1) | rs1059702 | F1(FITC) | AGGGGCCAGCAAAACGGxAA | 99 | 67.5 | 2.8 | |
F2(Texas Red) | AGGGGCCAGCAAAACGGxGA | ||||||
R(Biotin) | GGGACCCCACCTGATCCAGGC | ||||||
Melting curve method | |||||||
Gene | rs No | Primers | Sequence(5'→3') | Amplicon (bp) | Annealing temp.(°C) | Mg(mM) | Probe(5'→3') |
Tumor necrosis factor receptor superfamily member 1B (TNFRSF1B) | rs1061624 | F | CCC TCT GAC CTG CAG GCC AAG | 61 | 65 | 2 | AGC AGA GGC AGC GAG TTG |
R | CCA TGG CAG CAG AGG CTT TCC | ||||||
C-reactive protein 2 (CRP2) | rs1130864 | F | GAGCTCGTTAACTATGCTGGGA | 97 | 65 | 3 | GCTGGGAAACGGTCCAAA |
R | TTATCTCCAAGATCTGTCCAACTTG | ||||||