Insight into Gene Polymorphisms Involved in Toll-Like Receptor/Interferon Signalling Pathways for Systemic Lupus Erythematosus in South East Asia

Table 3

Touchdown PCR primers and conditions.

SNP	Primer sequence	𝑇𝑚	Final concentration	Inner/outer primers ratio	Mg^2+	Annealing temperature
STAT4
rs10168266	Forward inner primer (T allele) (29 bp); 5′-CAAAGTAGTAGCTATTGACTACATGAGAT	57°C	1.0 μM	4 : 1	2.5 mM	55°C
	Reverse inner primer (C allele) (27 bp); 5′-GTTATTACTACGGGTGGGTAGACATTG	62°C	1.0 μM
	Forward outer primer (28 bp); 5′-AAAAGTATAGAATTTGGAGGAAGAGAGT	59°C	0.25 μM
	Reverse outer primer (28 bp); 5′-TATTGGGGTATACTGAAAAGAAAGAGTA	59°C	0.25 μM
rs7601754	Forward inner primer (A allele) (21 bp); 5′-GGGTGAAGAAAAGGAACTCCA	60°C	1.0 μM	5 : 1	1.25 mM	55°C
	Reverse inner primer (G allele) (23 bp); 5′-CAAGGTCTTAGTATCATCTTGGC	57°C	1.0 μM
	Forward outer primer (28 bp); 5′-GGAGGTGATTACTATATTTCTAGGCTAA	58°C	0.2 μM
	Reverse outer primer (27 bp); 5′-AAAAATTAAAAATTAGTTGGCTATGGT	58°C	0.2 μM
TNFAIP3
rs2230936	Forward inner primer (G allele) (28 bp); 5′-CAGACTTGGTACTGAGGAAGGCGCTATG	69°C	1.0 μM	4 : 1	2.5 mM	62°C
	Reverse inner primer (T allele) (23 bp); 5′-GTCTGTTTCCTTGAGCGTGCCGA	69°C	1.0 μM
	Forward outer primer (28 bp); 5′-CTGAAAACCTTTGCTGGGTCTTACATGC	69°C	0.25 μM
	Reverse outer primer (29 bp); 5′-GACCTAGTCCATCAGATGCTACCAGAGGG	69°C	0.25 μM
rs3757173	Forward inner primer (T allele) (26 bp); 5′-GACCTTATTCCCTTCCCTGAAATGAT	64°C	1.0 μM	4 : 1	2.5 mM	53°C
	Reverse inner primer (C allele) (27 bp); 5′-CCTTAGCTGCAGACTAAGGTGGTATTG	64°C	1.0 μM
	Forward outer primer (28 bp); 5′-TTAAACCATTCAGTCCCCTAGAATAGCA	64°C	0.25 μM
	Reverse outer primer (28 bp); 5′-TAAAATCTTCCTACTGCCCATCTCTTTC	64°C	0.25 μM
IRF5
rs4728142	Forward inner primer (A allele) (26 bp); 5′-GTCACACCCCAAAAAGCTCTGAGACA	68°C	2.0 μM	5 : 1	1.25 mM	55°C
	Reverse inner primer (G allele) (26 bp); 5′-CCTTCCTCCCCATTTCTTACTAACCCC	68°C	2.0 μM
	Forward outer primer (28 bp); 5′-GAAAGGTGGAGACTCCGAGTGTAGAGGT	68°C	0.2 μM
	Reverse outer primer (28 bp); 5′-GACAGAGCGATACTCCGTCTCAAAAGAA	68°C	0.2 μM