Genetic polymorphism in CD14 gene, a co-receptor of TLR4 associated with non-alcoholic fatty liver disease

Table 1 Primer sequences, polymerase chain reaction conditions, and product sizes of different genes

Polymorphism site	Primer sequence (5'-3')	PCR conditions	Product size (bp)
TLR4 (Asp299gly)		(1) 95 °C for 10 min	249
		(2) 35 cycles of :
	F: GATTAGCATACTTAGACTACTACCTCCATG	94 °C for 1 min
	R: GATCAACTTCTGAAAAAGCATTCCCAC	61.5 °C for 1 min
		72 °C for 1 min
		(3) 72 °C for 7 min
TLR4 (Thr399Ile)		(1) 95 °C for 10 min	406
		(2) 35 cycles of :
	F: GGTTGCTGTTCTCAAAGTGATTTTGGGAGAA	94 °C for 1 min
	R: CCTGAAGACTGGAGAGTGAGTTAAATGCT	64.8 °C for 1 min
		72 °C for 1 min
		(3) 72 °C for 10 min
TLR2 (Arg753Gln)		(1) 95 °C for 10 min	289
		(2) 35 cycles of :
	F: 5’-CCTTCAAGTTGTGTCTTCATAAG-3’	94 °C for 1 min
	R: 5’-GGCCACTCCAGGTAGGTCTT-3’	58.6 °C for 1 min
		72 °C for 1 min
		(3) 72 °C for 10 min
CD14 (-550C/T)		(1) 95 °C for 10 min	368
		(2) 32 cycles of :
	F: GGAAGGGGGAATTTTTCTTTAGGC	94 °C for 1 min
	R: -GGCAGTGTCCTGATGACTCA	59.8 °C for 1 min
		72 °C for 1 min
		(3) 72 °C for 7 min
CD14 (-159C/T)		(1) 95 °C for 10 min	296
		(2) 35 cycles of :
	F: ATCATCCTTTTCCCACACC	94 °C for 40 s
	R: AACTCTTCGGCTGCCTCT	61 °C for 40 s
		72 °C for 40 s
		(3) 72 °C for 10 min