Table 1. PCR conditions of rs1256049, rs4986938, rs1256030 polymorphisms in the ESR2 gene.

	rs1256049 (+1082G/A, exon 5)	rs4986938 (+1730G/A, 3′UTR, exon 8)	rs1256030T/C (Intron 2)
*PCR Primers: sense antisense	5′-TTGCGCAGCTTAACTTCAAA-3′ 5′-ACCTGTCCAGAACAAGATCT-3′	5′-CAATGCATATCCTGCCTGTG-3′ 5′-GGTTTAGGGGTGGGGTAGACTG-3′	5′-CAATGCATATCCTGCCTGTG-3′ 5′-TCCCGGAAATCTGATACAGC-3′
PCR product size	321 bp	442 bp	251 bp
PCR conditions	Buffer enzyme 1X, 7.5 pmol of each primer, 0.2 mM of dNTPs, 2.0 mM of MgCl2, 2.5 u of Taq polymerase, 100 nM of genomic DNA	Buffer enzyme 1X, 7.5 pmol of each primer, 0.2 mM of dNTPs, 2.0 mM of MgCl2, 2.5 u of Taq polymerase, 100 nM of genomic DNA	Buffer enzyme 1X, 7.5 pmol of each primer, 0.2 mM of dNTPs, 2.5 mM of MgCl2, 2.5 u of Taq polymerase, 100 nM of genomic DNA
Annealing PCR temperature	60 °C	55 °C	57 °C
Recognition enzyme restriction	RsaI, 37 °C, (5′…GT↓AC…3′)	AluI, 37 °C, (5′…AG↓CT…3′)	AluI, 37 °C, (5′…AG↓CT…3′)
**Allele identified: (Wild type) (Polymorphic type)	G: 321 A: 211 + 110	G: 442 A: 336 + 106	C: 227 + 24 T: 147 + 80 + 24

Notes:

^*The primers previously described (Mahdavipour et al., 2017).

^**The genotypes were identified in 6% polyacrylamide gels (29:1), followed by silver nitrate staining.