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Determination of an optimal set/number of internal control  microRNA (miRNA) genes is a critical, but often undervalued, detail of  quantitative gene expression analysis. No validated internal genes for miRNA  quantitative PCR (q-PCR) in pig milk were available. We compared the  expression stability of six porcine internal control miRNA genes in pig milk  from different lactation periods (1 h, 3 days, 7 days, 14 days, 21 days, and 28  days postpartum), using an EvaGreen q-PCR approach. We found that using the  three most stable internal control genes to calculate the normalization factor  is sufficient for producing reliable q-PCR expression data. We also found that  miRNAs are superior to ribosomal RNA (rRNA) and snRNA, which are commonly used  as internal controls for normalizing miRNA q-PCR data. In terms of economic and  experimental feasibility, we recommend the use of the three most stable  internal control miRNA genes (miR-17, -107 and -103) for  calculating the normalization factors for pig milk samples from different  lactation periods. These results can be applied to future studies aimed at  measuring miRNA abundance in porcine milk.

Short Communication Suitable internal control microRNA genes for measuring miRNA abundance in pig milk during different lactation periods