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. 2020 Jul 15;82(9):1253–1259. doi: 10.1292/jvms.20-0301

Table 2. Summary of the primers, polymerase chain reaction (PCR) conditions, and genotyping methods used for the four single-nucleotide polymorphisms.

Gene Polymorphism Primer pairs PCR conditions Restriction enzyme RFLP a) Genotype
IL4R rs9193906; G >C 5′-GGCTCAGACCTACAACAGCA-3′ 94°C, 30 sec; 57°C, 15 sec; 72°C, 25 sec for 30 cycles PmlI C: 259 bp (uncut); G: 210 & 49 bp
5′-GCTCATTTGTTACCGCAGCC-3′
rs24378020; G >A 5′-CCCCTGTTCACCTTTGGACT-3′ 94°C, 30 sec, 60°C, 5 sec; 72°C, 10 sec for 30 cycles PvuII G: 309 bp (uncut); A: 191 & 118 bp
5′-TGAGGACCTGTCTCCACAGC-3′
rs851400460; G >A 5′-GTGGCAAGGCCCACATAGT-3′ 94°C, 30 sec; 60°C, 10 sec; 72°C, 20 sec for 30 cycles AvaI A: 311 bp (uncut); G: 160 & 151 bp
5′-CCTGCAGACATCAGCAACCA-3′

IL13 rs22147008; A >G 5′-TCTCAAACCCCACCTCCTGT-3′ 94°C, 30 sec; 65°C, 15 sec; 72°C, 25 sec for 30 cycles PspOMI A: 230 bp (uncut); G:168 & 62 bp
5′-AGGACAGAGGGCCTTACCC-3′

a) RFLP: restriction fragment length polymorphism.