Table 1 Primers sequences, product size and annealing temperatures

F5:c.1601G > A (rs6025)

FI primer (A allele):

RI primer (G allele):

FO primer (5′ - 3′):

RO primer (5′ - 3′):

TAAGAGCAGATCCCTGGACAGTCA

ACTTCAAGGACAAAATACCTGTATTCATC

TTTGAATATATTTTCTTTCAGGCAGGAA

AAATGTTATCACACTGGTGCTAAAAAGG

Product size for A allele: 184

Product size for G allele: 254

Product size of two outer primers: 385

62 °C

F5:c.6665A > G (rs6027)

FI primer (A allele):

RI primer (G allele):

FO primer (5′ - 3′):

RO primer (5′ - 3′):

TCGCCTGGAACTCTTTGGCTGGGA

CAGGGGTTTTTGAATGTTCAATTCTAGTAAAGAC

GGATAAAAAATTTTCTGGGTTGGGCATGAT

AAGCCCTTTTCTTGTTGGTTCTTGATATTTTCC

Product size for A allele: 291

Product size for G allele: 402

Product size of two outer primers: 635

60 °C

MTHFR:c.665C > T (rs1801133)

FI primer (T allele):

RI primer (C allele):

FO primer (5′ - 3′):

RO primer (5′ - 3′):

AAGGAGAAGGTGTCTGCGGGCGT

CAAAGAAAAGCTGCGTGATGATGAAATAGG

CTGAGAGTCATCTCTGGGGTCAGAAGCA

GAAGAACTCAGCGAACTCAGCACTCCAC

Product size for T allele: 246

Product size for C allele: 314

Product size of two outer primers: 507

60 °C

MTHFR:c.1286A > C (rs1801131)

FI primer (A allele):

RI primer (C allele):

FO primer (5′ - 3′):

RO primer (5′ - 3′):

TGTGGGGGGAGGAGCTGACCAGTGAGGA

GGTAAAGAACGAAGACTTCAAAGACACCTG

GCAAATACATCTTTGTTCTTGGGAGCGGG

TACCCTTCTCCCTTTGCCATGTCCACAG

Product size for A allele: 230

Product size for C allele: 316

Product size of two outer primers: 488

60 °C

VEGFA: c.-2055A > C (rs699947)

FI primer (C allele):

RI primer (A allele):

FO primer (5′ - 3′):

RO primer (5′ - 3′):

AGCTGTAGGCCAGACCCTGGTAC

AGTCAGTCTGATTATCCACCCAGACCT

CTTAGGACACCATACCGATGGAACTG

GCATATAGGAAGCAGCTTGGAAAAATTC

Product size for C allele: 161

Product size for A allele: 230

Product size of two outer primers: 341

60 °C

VEGFA:c.*237C > T (rs3025039)

FI primer (T allele):

RI primer (C allele):

FO primer (5′ - 3′):

RO primer (5′ - 3′):

ATTCCCGGGCGGGTGACCCAGAAT

AATGGCGAATCCAATTCCAAGAGGGAACG

ATACTGGGGCTTTCTGCCCCAGGACCAC

TGAGTGGGAACATTCCCCTCCCAACTCA

Product size for T allele: 310

Product size for C allele: 458

Product size of two outer primers: 715

62 °C