Table 2.

Oligonucleotide Primers Used in Multiplex PCR Reactions

	Gene (size of PCR product)	To detect mutation(s)	Forward primer	Reverse primer
10× Primer mix 1
	Cx26 (286 bp)	W24X, 35delG, M34T, V37I	5′-TCTTTTCCAGAGCAAACCGC-3′	5′-GsAsCsAsCsGAAGATCAGCTGCA-3′
	MtDNA 12 S RNA (137 bp)	A1555G	5′-CGTCACCCTCCTCAAGTATACTTC-3′ (2 μmol/L)	5′-GsCsTsTsTsGTGTTAAGCTACACTCTGG-3′ (2 μmol/L)
10× Primer mix 2
	SLC26A4 (249 bp)	L236P	5′-GGTTTCTATCTCAGGCAAACAT-3′	5′-AsTsTsGsTsTTCTGGAATGAACAGTGACC-3′
	SLC26A4 (139 bp)	IVS8 + 1G>A	5′-TTCAGACGATAATTGCTACTG-3′	5′-GsAsCsTsGsACTTACTGACTTAATG-3′
	SLC26A4 (215 bp)	E384G, T416P	5′-GTAGGATCGTTGTCATCCAG-3′	5′-CsGsAsGsCsCTTCCTCTGTTGC-3′
10× Primer mix 3
	GJB2 (311 bp)	L90P, R143W, 167delT, 235delC, 313del14	5′-CTGCAGCTGATCTTCGTGTC-3′	5′-AsCsAsAsAsGCAGTCCACAGTGTT-3′
	USH2A (159 bp)	2299delG	5′-ATGTGAGCCCTGCCAGTGTA-3′	5′-TsCsAsCsAsGGCCTTACAATTGGTG-3′

The 10× primer mixes are 4 μmol/L of each primer unless indicated. s, phosphorothioate bond.