T-Cell Cytokine Gene Polymorphisms and Vitamin D Pathway Gene Polymorphisms in End-Stage Renal Disease due to Type 2 Diabetes Mellitus Nephropathy: Comparisons with Health Status and Other Main Causes of End-Stage Renal Disease

Table 1

HRM and RFLP conditions for the identification of polymorphisms genotyped in the vitamin D pathway related genes.

Gene symbol	rs number	Alleles	Primers for PCR amplification (5′-3′)	Annealing temp. (°C)	PCR product length (bp)	HRM^a analysis	RFLP^b analysis
						Melting temp. range (°C)	Restriction enzyme	Restriction fragment length (bp)
GC	rs7041	G/T	F: GGAGGTGAGTTTATGGAACAGC	66.3	493		HaeIII	T = 493
			R: GGCATTAAGCTGGTATGAGGTC					G = 414 + 79
	rs1155563	C/T	F: GGTTATTCTAAGACTGTGCTCTTGC	63.0	116	71–78
			R: ATGTGTTCTCACTGTTCGACTCC
	rs2298849	C/T	F: TCCACTGGCAAAACACATTAC	60.6	118	73–83
			R: GGGACATCTGCATTTATCCTG
RXRA	rs10881578	A/G	F: TCTTGAGCAATGCCAGCAG	60.6	75	80–90
			R: CCACAGCTCACACATCCAATC
	rs10776909	C/T	F: CAGCCTGTGGCCTGCTCA	60.6	95	82–92
			R: AACCTCCGGCCCTTGGAG
	rs749759	A/G	F: ATAGGGCTTGCCTGCCTAGA	62.6	382		BstXI	A = 382
			R: CTCCACCATAGCCCAAGTGA					G = 243 + 139
VDR	rs1544410	A/G	F: GGAGACACAGATAAGGAAATAC	60.6	248		FspI	A (B) = 248
			R: CCGCAAGAAACCTCAAATAACA					G (b) = 175 + 73
	rs2228570	C/T	F: GCACTGACTCTGGCTCTGAC	72.5	341		FokI	C (F) = 341
			R: ACCCTCCTGCTCCTGTGGCT					T (f) = 282 + 59

HRM analysis: high resolution melt analysis.
^bRFLP analysis: restriction fragment length polymorphism analysis.