Table 1

Primers and PCR conditions for PCR-SSCP, PCR-RFLP, and others

Region	Primers	Sequence (5′ → 3′)	Size/Location^1-a	Conditions 30 Cycles	SSCP Running Temperature
Exon 4	EX4(S)	GTGAAGTTTAATCAGCTCCGTTG	282 bp/7287/7578	45 s 94°C	10°C
	EX4(R)	GATTGCAAGATGTTACCACTG		45 s 59°C
				45 s 72°C
Exon 5	EX5(S)	CATGAAGATCACCACAACTAATGTG	252 bp/12883/13134	45 s 94°C	15°C
	EX5(P)	CTTGGAAACGGACTGTGATCTTAC		45 s 59°C
				45 s 72°C
Exon 6	EX6(S)	GGACTTGAGTCTGCTCATTTAACTAC	230 bp/13260/13489	45 s 94°C	10°C
	EX6(R)	AGTAACCCAACAGTGCGACTG		45 s 58°C
Exon 7	EX7(S)	GCTGCATGTATAGTGGAAGGAC	317 bp/14619/14935	45 s 94°C	10°C
	EX7(R)	GGAATTGTACCTTTTAAGTGGATG		45 s 59°C
				45 s 72°C
Exon 8	EX8(S)	CTGATGGTGGTTAAATGACGATGT	287 bp/15776/16062	45 s 94°C	10°C
	EX8(R)	AAAGGAATGGGGTTCCAG		45 s 58°C
				45 s 72°C
Exon 10	EX10(S)	TCATCTAAGCTGTGATGTTGTACG	356 bp/19277/19632	30 s 94°C	15°C
	EX10(R)	GGCTTCCTACATTATGTCAGTGAAG		30 s 58°C
				30 s 72°C
Exon 11	EX11(S)	CAATAGTACTGCATGGACTCAGTTGAG	363 bp/27146/27508	45 s 94°C	15°C
	EX11(R)	GGCAGAATATGCTTGAACCAGC		45 s 63°C
				45 s 72°C
Exon 12	EX12(S)	GAACCCCTAACATGTAACTCTGTGG	346 bp/29699/30044	30 s 94°C	10°C
	EX12(R)	GTAGGTTCTTTGGCCCATAGAATG		30 s 59°C
				45 s 72°C
CYP3A5 5′ flanking region^1-b	3A51F	GGAAGCAACCTACATGTCCATC	1338 bp/−1234/104	45 s 94°C
	3A52R	ATCGCCACTTGCCTTCTTCAAC		30 s 61°C
				150 s 72°C
	A⁻⁴⁴G(S)	CAGGTGAGAGGATATTTAAGAGGC	337 bp/−232/105	30 s 94°C
	A⁻⁴⁴G(R)	CATCGCCACTTGCCTTCTTCAAC		30 s 61°C
				45 s 72°C
CYP3A5 minigene	MS	CTGTGGACTATGGCTGTCCAAATGTTC	6478 bp/8458/14935	two step:
	MR	GGAATTGTACCTTTTAAGTGGATG		16 cycles
				15 s 94°C 8 min 58°C 12 cycles 15 s 94°C 8 min 58°C (15 s per cycle)
pcDNA3.1/V5/His-TOPO vector	pcDNA3.1S	ACGACTCACTATAGGGAG	(sequencing primers)

CYP3A5 cDNA	pcDNA3.1R	CGAGGCTGATCAGCGGGTTTA	503 bp/12982/17177	45 s 94°C
	XL-4kb(S)	GGATGAAGAATGGAAGAGAATACGG		30 s 58°C
	XL-4kb(R)	GGACTCAGTTTCTTTCGAATTCTGG		30 s 72°C

bp, base pair.
↵1-a The complete genomic DNA sequence of CYP3A5 is according to Genbank: AC005020 [Homo sapiens BAC clone GSI-259H13 from seven complete sequences. Location number is from 15810 (the transcriptional initiation site, +1) to 47610 (exon 13)].
↵1-b Primer locations are based on CYP3A5 sequence data of Jounaidi et al. (1994).