Table 1. RT-PCR: primers, amplification,... | American Society of Hematology

Table 1.

RT-PCR: primers, amplification, and products

Gene	Sequence of primers, 5′ to 3′	Conditions of amplification	Product length, bp
VEGF₁₆₅	Forward: GCTGCACCCATGGCAGAAGG^*	30 cycles at 95°C 1 minute and 30 seconds, 95°C 1 minute, 65°C 30 seconds, 72°C 30 seconds	367
	Reverse: GAGCAAGGCCCACAGGGATTT†
SEMA3A	Forward: GGCATATAATCAGACTCACTTGTACGC	1 cycle at 94°C 10 minutes, 30 cycles at 94°C 30 seconds, 50°C 30 seconds, 72°C 30 seconds, and 1 cycle at 72°C 7 minutes	450
	Reverse: CTTGCATATCTGACCTATTCTAGCGTG
NRP1	Forward: GGCTCCAAATAGACCTGGGG	40 cycles at 95°C 1 minute and 30 seconds, 95°C 1 minute, 60°C 30 seconds, 72°C 30 seconds	600
	Reverse: GGTGCTGTCTATGACCGTGG
Plexin-A1	Forward: CCTCGAGAGCAAGAACCACCCCAAGCTGCT	1 cycle at 94°C 10 minutes, 30 cycles at 94°C 30 seconds, 68°C 30 seconds, 72°C 45 seconds, and 1 cycle at 72°C 7 minutes	150
	Reverse: CCCTTCACCGGCACCTCAGGTGCATT
GAPDH	Forward: CCCTCCAAAATCAAGTGGGG	20 cycles at 95°C 1 minute and 30 seconds, 95°C 45 seconds, 60°C 45 seconds, and 72°C 30 seconds	347
	Reverse: CGCCACAGTTTCCCGGAGGG

Gene	Sequence of primers, 5′ to 3′	Conditions of amplification	Product length, bp
VEGF₁₆₅	Forward: GCTGCACCCATGGCAGAAGG^*	30 cycles at 95°C 1 minute and 30 seconds, 95°C 1 minute, 65°C 30 seconds, 72°C 30 seconds	367
	Reverse: GAGCAAGGCCCACAGGGATTT†
SEMA3A	Forward: GGCATATAATCAGACTCACTTGTACGC	1 cycle at 94°C 10 minutes, 30 cycles at 94°C 30 seconds, 50°C 30 seconds, 72°C 30 seconds, and 1 cycle at 72°C 7 minutes	450
	Reverse: CTTGCATATCTGACCTATTCTAGCGTG
NRP1	Forward: GGCTCCAAATAGACCTGGGG	40 cycles at 95°C 1 minute and 30 seconds, 95°C 1 minute, 60°C 30 seconds, 72°C 30 seconds	600
	Reverse: GGTGCTGTCTATGACCGTGG
Plexin-A1	Forward: CCTCGAGAGCAAGAACCACCCCAAGCTGCT	1 cycle at 94°C 10 minutes, 30 cycles at 94°C 30 seconds, 68°C 30 seconds, 72°C 45 seconds, and 1 cycle at 72°C 7 minutes	150
	Reverse: CCCTTCACCGGCACCTCAGGTGCATT
GAPDH	Forward: CCCTCCAAAATCAAGTGGGG	20 cycles at 95°C 1 minute and 30 seconds, 95°C 45 seconds, 60°C 45 seconds, and 72°C 30 seconds	347
	Reverse: CGCCACAGTTTCCCGGAGGG

This primer is located in exon 2 (part of the conserved region of all VEGF splice variants)

This primer spans the boundaries of exons 5 and 7