Single Nucleotide Polymorphisms of the GJB2 and GJB6 Genes Are Associated with Autosomal Recessive Nonsyndromic Hearing Loss

Table 3

AS-PCR primers and conditions.

Gene	SNP ID; (nucleotide change)	Sequence (5′-3′)^*	Annealing temperature	Mg^2+	Amplicon size
GJB2	rs3751385; (C/T)	C allele: GCTCAGCTGTCAAGGCTCAGTCTCC; T allele: GCTCAGCTGTCAAGGCTCAGTCTCT; COM: TTGTCCTCAGAGAAAGAAACAAATGCC	60°C (1 min)	2.5 mM	284 bp
	rs7994748; (C/T)	C allele: GCTGAGAGCTGGGTTCCGTGTC; T allele: GCTGAGAGCTGGGTTCCGTGTT; COM: AGGGGCTCAGAAGCAGGACG	60°C (1 min)	2.5 mM	435 bp
	rs7329857; (C/T)	C allele: TTTCCCAACACAAAGATTCTGCC; T allele: TTTCCCAACACAAAGATTCTGCT; COM: CTTACACCAATAACCCCTAACAGCC	58°C (1 min)	2.5 mM	199 bp
	rs7987302; (G/A)	G allele: GGCATATCAGTCTATGGACAATGGGG; A allele: GGCATATCAGTCTATGGACAATGGGA; COM: AGAGGTTGCAGTGAGCCAAGG	58°C (1 min)	1.7 mM	157 bp
GJB6	rs7322538; (G/A)	G allele: CTAATGCAACTAGGGAAATTCG; A allele: CTAATGCAACTAGGGAAATTCA; COM: GCAATCTAGTTTTTCCTCATCC	56°C (45 s)	2.2 mM	106 bp
	rs9315400; (C/T)	C allele: GCAGCCTAGCATTTTACATC; T allele: GCAGCCTAGCATTTTACATT; COM: GTCTCTTTTTCGCAACCTTG	55°C (45 s)	2.5 mM	100 bp
	rs877098; (C/T)	C allele: AAGGGAGCTTGGAAATGAAGTC; T allele: AAGGGAGCTTGGAAATGAAGTT; COM: GAGGTGGAGCTTGCAGTGAG	56°C (1 min)	2.5 mM	227 bp
	rs945369; (A/C)	A allele: GTCCCTGTTTTTAGAAAAAAAGAA; C allele: GTCCCTGTTTTTAGAAAAAAAGAC; COM: GGAAGTAAACAGATCAGGGAG	59°C (1 min)	2.5 mM	187 bp
	rs7333214; (T/G)	T allele: AACATTTATCCAGGAATTGATATT; G allele: AACATTTATCCAGGAATTGATATG; COM: CAAATTTGCCAACAGACAATGC	57°C (1 min)	2.5 mM	230 bp
		Controls primers
AMELX		CTLA: CCCACCTTCCCCTCTCTCCAGGCAAATGGG; CTLB: GGGCCTCAGTCCCAACATGGCTAAGAGGTG			360 bp

COM: common primer (reverse). *The mismatches of the allele-specific primers are underlined. SNP ID: identification of the SNP. AMELX: human amelogenin gene used as an internal amplification control.