Table 1. Primers and conditions used for... | American Society of Hematology

Table 1.

Primers and conditions used for PCR analysis

Gene, PCR type	Direction	Primer	Product size, bp	Annealing temperature, °C	Cycle no.
β-actin
RT-PCR	F	GAC ATG GAG AAG ATC TGG CA	148	55	25
	R	GGG TGT TGA AGG TCT CAA AC
Brachyury
RT-PCR	F	CAT GTA CTC TTT CTT GCT GG	313	60	35
	R	GGT CTC GGG AAA GCA GTG GC
qPCR	F	CTC ACC AAC AAG CTC AAT GG	129	57	NA
	R	GGT CTC GGG AAA GCA GTG GC
*Fgf5*
RT-PCR	F	GTG TCT CAG GGG ATT GTA GGA ATA CG	314	65	35
	R	GTG AAG GAA AGT TCC GGT TGC
*Flk1*
RT-PCR + qPCR	F	CAC CTG GCA CTC TCC ACC TTC	219	55	35
	R	GAT TTC ATC CCA CTA CCG AAA G
*Gata1*
RT-PCR	F	CAT TGG CCC CTT GTG AGG CCA GAG A	289	60	35
	R	ACC TGA TGG AGC TTG AAA TAG AGG C
qPCR	F	CAA CAG TAT GGA GGG AAT TCC T	161	57	NA
	R	GTG TCC AAG AAC GTG TTG TTG C
*Gata2*
RT-PCR + qPCR	F	GCA CCT GTT GTG CAA ATT GT	195	57	35
	R	GCC CCT TTC TTG CTC TTC TT
*Gapdh*
qPCR	F	CAA TGA CCC CTT CAT TGA CC	145	55	35
	R	GAT CTC GCT CCT GGA AGA TG
*Klf1*
RT-PCR + qPCR	F	TCC ATC AGT ACA CTC ACC AC	92	57	35
	R	AAA TCC TGC GTC TCC TCA GA
*Klf6*
RT-PCR	F	TAT CTT CAG GAT GAG CCC TGC TAC	276	60	30
	R	AGA CTT CAC CAA TGG GAT CAG AGG
qPCR	F	ACC CGA CAT GGA TGT GCT CCC AAT	150	57	NA
	R	GCA GGG CTC ACT CTG AAG ATA
*Klf7*
RT-PCR	F	CAG CAT GGA CGT GTT GGC TAG TTA	276	60	33
	R	TCT TCT CGC AGA TGG TGG CTT CCA
*p21*
RT-PCR	F	GCA GAT CCA CAG CGA TAT CCA G	332	60	33
	R	GAG GAA GTA CTG GGC CTC TTG T
*Rex1*
RT-PCR + qPCR	F	CGT GTA ACA TAC ACC ATC CG	129	60	35
	R	GAA ATC CTC TTC CAG AAT GG
*Scl*
RT-PCR + qPCR	F	ATA GCC TTA GCC AGC CGC TC	153	55	33
	R	CGC CGC ACT ACT TTG GTG TG

Gene, PCR type	Direction	Primer	Product size, bp	Annealing temperature, °C	Cycle no.
β-actin
RT-PCR	F	GAC ATG GAG AAG ATC TGG CA	148	55	25
	R	GGG TGT TGA AGG TCT CAA AC
Brachyury
RT-PCR	F	CAT GTA CTC TTT CTT GCT GG	313	60	35
	R	GGT CTC GGG AAA GCA GTG GC
qPCR	F	CTC ACC AAC AAG CTC AAT GG	129	57	NA
	R	GGT CTC GGG AAA GCA GTG GC
*Fgf5*
RT-PCR	F	GTG TCT CAG GGG ATT GTA GGA ATA CG	314	65	35
	R	GTG AAG GAA AGT TCC GGT TGC
*Flk1*
RT-PCR + qPCR	F	CAC CTG GCA CTC TCC ACC TTC	219	55	35
	R	GAT TTC ATC CCA CTA CCG AAA G
*Gata1*
RT-PCR	F	CAT TGG CCC CTT GTG AGG CCA GAG A	289	60	35
	R	ACC TGA TGG AGC TTG AAA TAG AGG C
qPCR	F	CAA CAG TAT GGA GGG AAT TCC T	161	57	NA
	R	GTG TCC AAG AAC GTG TTG TTG C
*Gata2*
RT-PCR + qPCR	F	GCA CCT GTT GTG CAA ATT GT	195	57	35
	R	GCC CCT TTC TTG CTC TTC TT
*Gapdh*
qPCR	F	CAA TGA CCC CTT CAT TGA CC	145	55	35
	R	GAT CTC GCT CCT GGA AGA TG
*Klf1*
RT-PCR + qPCR	F	TCC ATC AGT ACA CTC ACC AC	92	57	35
	R	AAA TCC TGC GTC TCC TCA GA
*Klf6*
RT-PCR	F	TAT CTT CAG GAT GAG CCC TGC TAC	276	60	30
	R	AGA CTT CAC CAA TGG GAT CAG AGG
qPCR	F	ACC CGA CAT GGA TGT GCT CCC AAT	150	57	NA
	R	GCA GGG CTC ACT CTG AAG ATA
*Klf7*
RT-PCR	F	CAG CAT GGA CGT GTT GGC TAG TTA	276	60	33
	R	TCT TCT CGC AGA TGG TGG CTT CCA
*p21*
RT-PCR	F	GCA GAT CCA CAG CGA TAT CCA G	332	60	33
	R	GAG GAA GTA CTG GGC CTC TTG T
*Rex1*
RT-PCR + qPCR	F	CGT GTA ACA TAC ACC ATC CG	129	60	35
	R	GAA ATC CTC TTC CAG AAT GG
*Scl*
RT-PCR + qPCR	F	ATA GCC TTA GCC AGC CGC TC	153	55	33
	R	CGC CGC ACT ACT TTG GTG TG

qPCR indicates quantitative real-time PCR; F, forward; R, reverse; and NA, not applicable.