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CLC number: S823.9+1

On-line Access: 2015-06-08

Received: 2014-12-14

Revision Accepted: 2015-04-29

Crosschecked: 2015-05-20

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Citations:  Bibtex RefMan EndNote GB/T7714


Hong-yun Liu


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Journal of Zhejiang University SCIENCE B 2015 Vol.16 No.6 P.533-540


Comparative studies of two methods for miRNA isolation from milk whey

Author(s):  Xiao-lu Jin, Zi-hai Wei, Lan Liu, Hong-yun Liu, Jian-xin Liu

Affiliation(s):  Institute of Dairy Science, MoE Key Laboratory of Molecular Animal Nutrition, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China

Corresponding email(s):   hyliu@zju.edu.cn, liujx@zju.edu.cn

Key Words:  Method, Milk whey miRNA, Spike-in miRNA

Xiao-lu Jin, Zi-hai Wei, Lan Liu, Hong-yun Liu, Jian-xin Liu. Comparative studies of two methods for miRNA isolation from milk whey[J]. Journal of Zhejiang University Science B, 2015, 16(6): 533-540.

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T1 - Comparative studies of two methods for miRNA isolation from milk whey
A1 - Xiao-lu Jin
A1 - Zi-hai Wei
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J0 - Journal of Zhejiang University Science B
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PB - Zhejiang University Press & Springer
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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 (<200 nt) from milk whey, and their yields were quite similar. However, the quantification results show that the total miRNA recovery efficiency by the TM method is superior to that by the TP method. The TM method performed better than the TP for recovery of milk whey miRNA due to its consistency and good repeatability in endogenous and spike-in miRNA recovery. Additionally, quantitative recovery analysis of a spike-in miRNA may be more accurate to reflect the milk whey miRNA recovery efficiency than using traditional RNA quality analysis instruments (NanoDrop or Bioanalyzer 2100).




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