Table 1.

Primer pairs and conditions for PCR

Gene	Acc. no.	Forward primer sequence (5′–3′)	Reverse primer sequence (5′–3′)	Binding position	Length (bp)	Cycle no.	Reference
α1C	L29534	TGA GAC CGA GTC CGT CAA A	GAA AAT CAC CAG CCA GTA GAA GA	1333–1522	190	30	Grammer et al. 2000
α1D	M83566	GCA AGA TGA CGA GCC TGA G	ATG GTT ATG ATG GTT ATG ACA C	5164–5407	244	30
α1G	AF134986	ATG GCC ATG GAG CAC TAC C	CGA GGC GTT GAC CTC GAT T	4882–5100	219	30	Huang et al. 2000
α1H	AF051946	CAC TCA TTC TAC AAC TTC ATC	CTC TCC CGC TGC TTC GTC	1250–1368	190	30
α1S	L33798	GGT GGA GGC TGC GAT GGA	ATG GCT GTT GCT ATG GTT GCT	4989–5262	274	30	Barry, 2000
GAPDH	J02642	AAC AGC GAC ACC CAC TCC TC	GGA GGG GAG ATT CAG TGT GGT	869–1126	258	29
HCN1	AF064876	TTG TCG TCT TTA CTC ACT TTC	CTC CTG ATT GTT GAA AAC AC	1311–1491	181	30
HCN2	AJ012582	GCC TGA TCC GCT ACA TCC A	TGC GAA GGA GTA CAG TTC AC	1078–1304	227	30	Ludwig et al. 1999
HCN4	AJ132429	CGC CTC ATT CGA TAT ATT CAC	CGC GTA GGA GTA CTG CTT C	1743–1970	228	30	Ludwig et al. 1999
Kir2.1	L36069	GAC CTG GAG ACG GAC GAC	AGC CTG GAG TCT GTC AAA GTC	910–1302	393	30	Wang et al. 1998b
Kir2.2	U16861	TTG AGT AAA CAG GAC ATT GAC	CTG GTT GTG AAG GTC TAT G	1171–1560	390	30	Wang et al. 1998b
Kir2.3	S72503	TAT GGC ATG GGC AAG GAG	AGC TGC CTC CTC CTC CAT C	912–1274	363	30	Wang et al. 1998b
Kir3.1	U50964	TCC CCT TGA CCA ACT TGA ACT	ACG ACA TGA GAA GCA TTT CCT C	928–1301	374	30
Kir3.4	U52154	TTT TCC AAC AAC GCA GTC A	CAC AAC TTC AAA CTC TTC C	983–1285	303	30
Kv1.1	L02750	CCA TCA TTC CTT ATT TCA TCA C	CTC TTC CCC CTC AGT TTC TC	782–1269	488	30
Kv1.4	L02751	GAG AGA AGA GGA AGA CAG GGC	TGG GGT GCT GAA GTA TCA TTC	828–1073	246	34	Ohya et al. 1997
Kv1.5	M83254	CAT TGC CCT GCC TGT GCC	TGC TCC CGC TGA CCT TCC	1677–1834	158	34
Kv2.1	L02840	TAC TGG GGC ATC GAC GAG A	GAC TGG CCG AAC TCA TCG A	547–854	308	34	Schultz et al. 2001
Kv3.1	S56770	AGG ACG AGC TGG AGA TGA CC	AAG AAG AGG GAA GCG AAG G	505–664	160	35	Wulfsen et al. 2000
Kv4.2	AJ010969	ATC TTC CGC CAC ATC CTG AA	GAT CCG CAC GGC ACT GTT TC	700–1061	362	34	Postma et al. 2000
Kv4.3	AF205857	GAT GAG CAG ATG TTT GAG CAG	AGC AGG TGG TAG TGA GGC C	1534–1639	106	28
Kv7.1	U89364	CAT CAT CGA CCT CAT CGT GG	TTC TCG GCA GCA TAG CAC CT	399–959	561	30	Lai et al. 1999
Kv7.2	AF033348	GGA CTC GCT TTC AGG AAG G	CCC TTC CCC TTG GCA G	1331–1494	164	35
Kv7.3	AF033347	GGA GAG GAG ATG AAA GAG GAG	TGA AGA AAG GAA AAG AGA CGA C	736–1093	358	35
MaxiK	U11058	ACA ACA TCT CCC CCA ACC	TCA TCA CCT TCT TTC CAA TTC	1222–1531	310	35
SCN5A	M77235	CCT AAT CAT CTT CCG CAT CC	TGT TCA TCT CTC TGT CCT CAT C	2814–3022	208	35
Twik1	U336321	TCC TGC TTC TTC TTC ATC	AGG CTC ATT TTG CTT CTG GTC	759–1143	385	30	Wang et al. 1998b

The table specifies forward and reverse primers used for semi-quantitative RT-PCR of various ion channels and the primers for the housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Primers were constructed with HUSAR program package (Senger et al. 1998) or modified using published sequences. Annealing temperature (T_A) was 60°C for all primers except for Kv7.2 (54°C) and Kv7.3 (58°C). After initial denaturation at 94°C for 5 min, cycling conditions were 94°C for 30 s, T_A for 30 s and 72°C for 30 s for all primers except for Kv7.1 (94°C for 60 s, T_A for 60 s and 72°C for 60 s) and Kv7.3 (94°C for 30 s, T_A for 30 s and 72°C for 60 s). Numbers of cycles are indicated for each primer pair. For a final extension reaction mixes were heated at 72°C for 7 min.