Primer sequences utilized for real time PCR analysis of genes involved in estrogen signaling in bovine mammary epithelial (MAC-T) cell line.

All primer sequences were designed to span exon-exon junctions, to minimize the potential of amplifying genomic DNA, using Primer3 software with sequences obtained from GenBank (<a href="http://www.ncbi.nlm.nih.gov/" target="_blank">http://www.ncbi.nlm.nih.gov/</a>). All primer pairs displayed melting curves with a single peak, indicative of a pure, single amplicon, confirmed the specificity of the primers. All efficiencies (10 ^(-1/slop) – 1 x 100]) ranged between 85% and 110%, with an R² value > 0.94. Last, primer binding specificity was tested in silico against the target genome or transcriptome to avoid off-target amplification. (DOCX)