Comparative studies of two methods for miRNA isolation from milk whey

• Published in: Journal of Zhejiang University. B. Science 2015-06, Vol.16 (6), p.533-540
• Main Authors: Jin, Xiao-lu, Wei, Zi-hai, Liu, Lan, Liu, Hong-yun, Liu, Jian-xin
• Format: Article
• Language: English
• Subjects: Animals; Biomedical and Life Sciences; Biomedicine; Fractional Precipitation - methods; Guanidines - chemistry; MicroRNAs - genetics; MicroRNAs - isolation & purification; Phenols - chemistry; Whey - chemistry
• ISSN: 1673-1581; 1862-1783
• DOI: 10.1631/jzus.B1400355

MicroRNAs (miRNAs) from milk whey have been considered for their potential as noninvasive biomarkers for milk quality control and disease diagnosis. However, standard protocols for miRNA isolation and quantification from milk whey are not well established. The objective of this study was to compare two methods for the isolation of miRNAs from milk whey. These two methods were modified phenol-based technique (Trizol LS® followed by phenol precipitation, the TP method) and combined phenol and column-based approach (Trizol LS® followed by cleanup using the miRNeasy kit, the TM method). Yield and quality of RNA were rigorously measured using a NanoDrop ND-1000 spectrophotometer and then the distribution of RNA was precisely detected in a Bioanalyzer 2100 instrument by microchip gel electrophoresis. Several endogenous miRNAs (bta-miR-141, bta-miR-146a, bta-miR-148a, bta-miR-200c, bta-miR-362, and bta-miR-375) and an exogenous spike-in synthetic control miRNA (cel-miR-39) were quantified by real-time polymerase chain reaction (PCR) to examine the apparent recovery efficiency of milk whey miRNAs. Both methods could successfully isolate sufficient small RNA (