Table 1. Oligonucleotide Primers Used... | American Society of Hematology

Table 1.

Oligonucleotide Primers Used for RT-PCR

Gene	Size (bp)^*	T (°C)^†	5′ Primer	3′ Primer	Internal Probe	Reference
Mesodermal markers
Brachyury	947	52	TGCTGCCTGTGAGTCATAAC	TCCAGGTGCTATATATTGCC	GAGCTCAGTTCTTTCGAGGC	49
BMP RIA	451	55	CCAGAAGTGCTGGATGAAAGCC	CCTATTCCATGCTAATCCCACC	ACAGCGATGAATGTCTTCGAG	6
BMP-4	570	55	ACTGTGAGGAGTTTCCATCACG	TCTTATTCTTCTTCCTGGACCG	GGACACCAGACTAGTCCATCAC	9
Smad1	336	55	CCAGGCGGCATATTGGAAAAGG	ATCTCAATCCAGCAGGGGGTGC	AGCCGGAACTGCAACTACCACC	69
						Transcription factors
scl	396	55	TATGAGATGGAGATTTCTGATG	GCTCCTCTGTGTAACTGTCC	GTCCTCACACCAAAGTAGTG	70
GATA-1	404	60	GGAATTCGGGCCCCTTGTGAGG	CGGGGTACCTCACGCTCCAG	GTCTGCAATGCCTGTGGCCTCTA	71
			CCAGAGAG	CCAGATTCGACCC
GATA-2	∼720	60	CGGAATTCGACACACCACCCGA	CGGAATTCGCCTACGCCATGGC	GTCTGCAATGCCTGTGGCCTCTA	59
			TACCCACCTAT	AGTCACCATGCT
EKLF	359	55	TCGCCGGAGACGCAGGCT	CCCAGTCCTTGTGCAGGA	ACGCAGCCGGCGAACTTTGG	49
NF-E2	391	62.5	GAGCCCTGGCCATGAAGATTCC	CACCATCAGCAGCCTGTTGCAG	GCAAGAACAAGGTGGCAGCCC	55
c-myb	681	55	CCTCTAGGAGCTCATTTGTG	TTCAAGGCCAGCATTCTTGC	TACTAAACCATTTCATGAGG	49
PU.1	615	55	TGGAAGGGTTTTCCCTCACC	TGCTGTCCTTCATGTCGCCG	GAGAACTTCCCTGAGAACCAC	72
						Receptors
CD34	618	55	GTTACCTCTGGGATCCCTTCAG	CTCCAGAGGTGACCAATGAATA	CAACCACAGACTTCCCCAAC	45
			GCTC	AG
flk-1	398	60	TAGGTGCCTCCCCATACCCTGG	TGGCCGGCTCTTTCGCTTACTG	GGCAAAGACTATATTGTTCTTC	73
flk-2	300	60	CTTCTGTGCAAATGCTGTGCGC	GTTCTCTCTCGGGAACTCCCAC	TTCTCATTGTGTTGATCTGCC	45
			GTAC	TTAAG
c-kit	765	50	TGTCTCTCCAGTTTCCCTGC	TTCAGGGACTCATGGGCTCA	ACAACACTCTTATCGTAGACC	49
Epo R	356	62	TTGACGCTGTCTCTCATTCTGG	CAATACCAAGTAGGTGTCCTGG	CGAGTTTGAGGGTCTCTTCACC	74
						Erythroid and myeloid genes
ζ globin	371	55	GCTCAGGCCGAGCCCATTGG	TAGCGGTACTTCTCAGTCAG	CGGTCAACTTCAAGCTCCTG	59
α globin	334	55	CTCTCTGGGGAAGACAAAAGCA	GGTGGCTAGCCAAGGTCACCAG	GCCCTGGAAAGGATGTTTGCTA	59
			AC	CA	GC
βH1 globin	265	55	CTCAAGGAGACCTTTGCTCA	AGTCCCCATGGAGTCAAAGA	TCCAATCACCAGCTTCTGCC	49
ε^y2 globin	157	55	GGAGAGTCCATTAAGAACCTAG	CTGTGAATTCATTGCCGAA	CCTGAGAACTTCAAACTCTTGG	59
			ACAA	GTAC	GT
β_maj globin	578	55	CACAACCCCAGAAACAGACA	CTGACAGATGCTCTCTTGGG	ACCTTCTGGAAGGCAGCCTGTGC	49
MPO	379	55	GGTGCTGAAGAACCTGGAGTTG	CTAGGTCTCCTTCCAGGAAGTC	GATGGTGATCGGTTTTGGTGG	75
Lysozyme	344	60	TGCAGGATGACATCACTGCAGC	GCTGTGCTGACTGACAAGGGAG	CATTCGAGCATGGGTGGCATG	76
vav	604	55	GTATAACGTGGAGGTCAAGCA	GGAGGACTGGAGAAAATCAGT	GCAGAATTCCCTCAAAGATTGC	49
LMO2	505	60	TCGCGAATTCCACCATGTCCTC	CGAGTTCTAGACTAGATGATCC	GAGGATTGCCTCAGCTGTGACC	29
			GGCCATCGAAAGGA	CATTGATCTTGGT
HPRT	249	55	GCTGGTGAAAAGGACCTCT	CACAGGACTAGAACACCTGC	GGATACAGGCCAGACTTTGT	49

Gene	Size (bp)^*	T (°C)^†	5′ Primer	3′ Primer	Internal Probe	Reference
Mesodermal markers
Brachyury	947	52	TGCTGCCTGTGAGTCATAAC	TCCAGGTGCTATATATTGCC	GAGCTCAGTTCTTTCGAGGC	49
BMP RIA	451	55	CCAGAAGTGCTGGATGAAAGCC	CCTATTCCATGCTAATCCCACC	ACAGCGATGAATGTCTTCGAG	6
BMP-4	570	55	ACTGTGAGGAGTTTCCATCACG	TCTTATTCTTCTTCCTGGACCG	GGACACCAGACTAGTCCATCAC	9
Smad1	336	55	CCAGGCGGCATATTGGAAAAGG	ATCTCAATCCAGCAGGGGGTGC	AGCCGGAACTGCAACTACCACC	69
						Transcription factors
scl	396	55	TATGAGATGGAGATTTCTGATG	GCTCCTCTGTGTAACTGTCC	GTCCTCACACCAAAGTAGTG	70
GATA-1	404	60	GGAATTCGGGCCCCTTGTGAGG	CGGGGTACCTCACGCTCCAG	GTCTGCAATGCCTGTGGCCTCTA	71
			CCAGAGAG	CCAGATTCGACCC
GATA-2	∼720	60	CGGAATTCGACACACCACCCGA	CGGAATTCGCCTACGCCATGGC	GTCTGCAATGCCTGTGGCCTCTA	59
			TACCCACCTAT	AGTCACCATGCT
EKLF	359	55	TCGCCGGAGACGCAGGCT	CCCAGTCCTTGTGCAGGA	ACGCAGCCGGCGAACTTTGG	49
NF-E2	391	62.5	GAGCCCTGGCCATGAAGATTCC	CACCATCAGCAGCCTGTTGCAG	GCAAGAACAAGGTGGCAGCCC	55
c-myb	681	55	CCTCTAGGAGCTCATTTGTG	TTCAAGGCCAGCATTCTTGC	TACTAAACCATTTCATGAGG	49
PU.1	615	55	TGGAAGGGTTTTCCCTCACC	TGCTGTCCTTCATGTCGCCG	GAGAACTTCCCTGAGAACCAC	72
						Receptors
CD34	618	55	GTTACCTCTGGGATCCCTTCAG	CTCCAGAGGTGACCAATGAATA	CAACCACAGACTTCCCCAAC	45
			GCTC	AG
flk-1	398	60	TAGGTGCCTCCCCATACCCTGG	TGGCCGGCTCTTTCGCTTACTG	GGCAAAGACTATATTGTTCTTC	73
flk-2	300	60	CTTCTGTGCAAATGCTGTGCGC	GTTCTCTCTCGGGAACTCCCAC	TTCTCATTGTGTTGATCTGCC	45
			GTAC	TTAAG
c-kit	765	50	TGTCTCTCCAGTTTCCCTGC	TTCAGGGACTCATGGGCTCA	ACAACACTCTTATCGTAGACC	49
Epo R	356	62	TTGACGCTGTCTCTCATTCTGG	CAATACCAAGTAGGTGTCCTGG	CGAGTTTGAGGGTCTCTTCACC	74
						Erythroid and myeloid genes
ζ globin	371	55	GCTCAGGCCGAGCCCATTGG	TAGCGGTACTTCTCAGTCAG	CGGTCAACTTCAAGCTCCTG	59
α globin	334	55	CTCTCTGGGGAAGACAAAAGCA	GGTGGCTAGCCAAGGTCACCAG	GCCCTGGAAAGGATGTTTGCTA	59
			AC	CA	GC
βH1 globin	265	55	CTCAAGGAGACCTTTGCTCA	AGTCCCCATGGAGTCAAAGA	TCCAATCACCAGCTTCTGCC	49
ε^y2 globin	157	55	GGAGAGTCCATTAAGAACCTAG	CTGTGAATTCATTGCCGAA	CCTGAGAACTTCAAACTCTTGG	59
			ACAA	GTAC	GT
β_maj globin	578	55	CACAACCCCAGAAACAGACA	CTGACAGATGCTCTCTTGGG	ACCTTCTGGAAGGCAGCCTGTGC	49
MPO	379	55	GGTGCTGAAGAACCTGGAGTTG	CTAGGTCTCCTTCCAGGAAGTC	GATGGTGATCGGTTTTGGTGG	75
Lysozyme	344	60	TGCAGGATGACATCACTGCAGC	GCTGTGCTGACTGACAAGGGAG	CATTCGAGCATGGGTGGCATG	76
vav	604	55	GTATAACGTGGAGGTCAAGCA	GGAGGACTGGAGAAAATCAGT	GCAGAATTCCCTCAAAGATTGC	49
LMO2	505	60	TCGCGAATTCCACCATGTCCTC	CGAGTTCTAGACTAGATGATCC	GAGGATTGCCTCAGCTGTGACC	29
			GGCCATCGAAAGGA	CATTGATCTTGGT
HPRT	249	55	GCTGGTGAAAAGGACCTCT	CACAGGACTAGAACACCTGC	GGATACAGGCCAGACTTTGT	49

Abbreviations: BMP, bone morphogenetic protein; Epo R, erythropoietin receptor; MPO, myeloperoxidase; HPRT, hypoxanthine phosphoribosyl transferase.

Size of the predicted PCR product.

†

Annealing temperature.

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