Table 2. RT-PCR Analysis of RNA... | American Society of Hematology

Table 2.

RT-PCR Analysis of RNA Transcripts: Primer Sequences, PCR Conditions, and Sizes of PCR Products

Gene	Primers	Sequence 5′→3′	Exon	Hybridization T°	Cycles No.	[MgCl₂] mmol/L	Size of PCR Product	Enzymes
IDS	IDS 3a	TGTACGACTTCAACT	3	47	35	2.5	158	HpaII
		CCTACTGGA
	(sense)
	IDS	AGACCAGCTATACG	4
		GAGAATCAC*C
	(antisense)
P55	P55 H	AGAGCCCATGGGAA	3-4	51	35	1.5	138	Hha I
		TCG*C
	(sense)
	P55 I	ACATTTGTGCCATTG	4
		ATTTCTAGG
	(antisense)
G6PD	G6PD-E10	GCTGGACCTGACCTA	10	59	35	3.0	79	Pvu I
		CGGCAACA
	(sense)
	G6PVPDPVU1	GAAGACGTCCAGGA	11
		TGAGGCGA*TC
	(antisense)

Gene	Primers	Sequence 5′→3′	Exon	Hybridization T°	Cycles No.	[MgCl₂] mmol/L	Size of PCR Product	Enzymes
IDS	IDS 3a	TGTACGACTTCAACT	3	47	35	2.5	158	HpaII
		CCTACTGGA
	(sense)
	IDS	AGACCAGCTATACG	4
		GAGAATCAC*C
	(antisense)
P55	P55 H	AGAGCCCATGGGAA	3-4	51	35	1.5	138	Hha I
		TCG*C
	(sense)
	P55 I	ACATTTGTGCCATTG	4
		ATTTCTAGG
	(antisense)
G6PD	G6PD-E10	GCTGGACCTGACCTA	10	59	35	3.0	79	Pvu I
		CGGCAACA
	(sense)
	G6PVPDPVU1	GAAGACGTCCAGGA	11
		TGAGGCGA*TC
	(antisense)

Indicate the mismatched nucleotides introduced by the primer to create restriction sites.