Table 3.

Primer sequences and conditions for pyrosequencing analysis

Gene	Primer sequence	Primer concentration [nM]	Ta [°C]	Amplicon length [bp]	No. of CpGs analyzed	Chromosome, position of sequence to analyze, DNA strand	Reference
ESR2	f: 5′ GGAGGTTGAGAGAAATAATTGTTTTTTGA 3′	400	61	255	9	14, [49,292- 49,369], positive strand	in-house [NG_011535.1]
	r: 5′ [Btn]ATAAACACACCCACCTTACCTTCTCTA 3′ s: 5′ GAAATAATTGTTTTTTGAAATTTG 3′

f forward primer, r reverse primer, s sequencing primer, [Btn] biotin, T _a annealing temperature