Full Text:   <2562>

Summary:  <1944>

CLC number: S823.9+1

On-line Access: 2015-06-08

Received: 2014-12-14

Revision Accepted: 2015-04-29

Crosschecked: 2015-05-20

Cited: 0

Clicked: 4003

Citations:  Bibtex RefMan EndNote GB/T7714


Hong-yun Liu


-   Go to

Article info.
Open peer comments

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.

@article{title="Comparative studies of two methods for miRNA isolation from milk whey",
author="Xiao-lu Jin, Zi-hai Wei, Lan Liu, Hong-yun Liu, Jian-xin Liu",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Comparative studies of two methods for miRNA isolation from milk whey
%A Xiao-lu Jin
%A Zi-hai Wei
%A Lan Liu
%A Hong-yun Liu
%A Jian-xin Liu
%J Journal of Zhejiang University SCIENCE B
%V 16
%N 6
%P 533-540
%@ 1673-1581
%D 2015
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1400355

T1 - Comparative studies of two methods for miRNA isolation from milk whey
A1 - Xiao-lu Jin
A1 - Zi-hai Wei
A1 - Lan Liu
A1 - Hong-yun Liu
A1 - Jian-xin Liu
J0 - Journal of Zhejiang University Science B
VL - 16
IS - 6
SP - 533
EP - 540
%@ 1673-1581
Y1 - 2015
PB - Zhejiang University Press & Springer
ER -
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).




Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


[1]Baier, S.R., Nguyen, C., Xie, F., et al., 2014. MicroRNAs are absorbed in biologically meaningful amounts from nutritionally relevant doses of cow milk and affect gene expression in peripheral blood mononuclear cells, HEK-293 kidney cell cultures, and mouse livers. J. Nutr., 144(10):1495-1500.

[2]Bartel, D.P., 2004. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell, 116(2):281-297.

[3]Chen, X., Gao, C., Li, H., et al., 2010. Identification and characterization of microRNAs in raw milk during different periods of lactation, commercial fluid, and powdered milk products. Cell Res., 20(10):1128-1137.

[4]Duy, J., Koehler, J.W., Honko, A.N., et al., 2015. Optimized microRNA purification from TRIzol-treated plasma. BMC Genomics, 16(1):95.

[5]Farazi, T.A., Horlings, H.M., Jelle, J., et al., 2011. MicroRNA sequence and expression analysis in breast tumors by deep sequencing. Cancer Res., 71(13):4443-4453.

[6]Fernández-Hernando, C., Suárez, Y., Rayner, K.J., et al., 2011. MicroRNAs in lipid metabolism. Curr. Opin. Lipidol., 22(2):86.

[7]Fleige, S., Pfaffl, M.W., 2006. RNA integrity and the effect on the real-time qRT-PCR performance. Mol. Aspects Med., 27(2-3):126-139.

[8]Guo, L., Lu, Z., 2010. The fate of miRNA* strand through evolutionary analysis: implication for degradation as merely carrier strand or potential regulatory molecule. PLoS ONE, 5(6):e11387.

[9]Hata, T., Murakami, K., Nakatani, H., et al., 2010. Isolation of bovine milk-derived microvesicles carrying mRNAs and microRNAs. Biochem. Biophys. Res. Commun., 396(2):528-533.

[10]Izumi, H., Kosaka, N., Shimizu, T., et al., 2012. Bovine milk contains microRNA and messenger RNA that are stable under degradative conditions. J. Dairy Sci., 95(9):4831-4841.

[11]Izumi, H., Kosaka, N., Shimizu, T., et al., 2013. Purification of RNA from milk whey. Methods Mol. Biol., 1024:191-201.

[12]Izumi, H., Kosaka, N., Shimizu, T., et al., 2014. Time-dependent expression profiles of microRNAs and mRNAs in rat milk whey. PLoS ONE, 9(2):e88843.

[13]Klein, M.S., Buttchereit, N., Miemczyk, S.P., et al., 2012. NMR metabolomic analysis of dairy cows reveals milk glycerophosphocholine to phosphocholine ratio as prognostic biomarker for risk of ketosis. J. Proteome Res., 11(2):1373-1381.

[14]Kosaka, N., Izumi, H., Sekine, K., et al., 2010. microRNA as a new immune-regulatory agent in breast milk. Silence, 1(1):7.

[15]Kroh, E.M., Parkin, R.K., Mitchell, P.S., et al., 2010. Analysis of circulating microRNA biomarkers in plasma and serum using quantitative reverse transcription-PCR (qRT-PCR). Methods, 50(4):298-301.

[16]Li, Y., Kowdley, K.V., 2012. Method for microRNA isolation from clinical serum samples. Anal. Biochem., 431(1):69-75.

[17]McAlexander, M.A., Phillips, M.J., Witwer, K.W., 2013. Comparison of methods for miRNA extraction from plasma and quantitative recovery of RNA from cerebrospinal fluid. Front. Genet., 4:83

[18]Melnik, B.C., John, S.M., Schmitz, G., 2013. Milk is not just food but most likely a genetic transfection system activating mTORC1 signaling for postnatal growth. Nutr. J., 12(1):103.

[19]Moret, I., Sánchez-Izquierdo, D., Iborra, M., et al., 2013. Assessing an improved protocol for plasma microRNA extraction. PLoS ONE, 8(12):e82753.

[20]Munch, E.M., Harris, R.A., Mohammad, M., et al., 2013. Transcriptome profiling of microRNA by Next-Gen deep sequencing reveals known and novel miRNA species in the lipid fraction of human breast milk. PLoS ONE, 8(2):e50564.

[21]Saleh, M., Afify, A.M., Kamel, A., 1998. Mother’s milk protein profile, a possible biomarker for human exposure to persistent insecticides. J. Environ. Sci. Health B, 33(6):645-655.

[22]Shi, R., Chiang, V.L., 2005. Facile means for quantifying microRNA expression by real-time PCR. Biotechniques, 39(4):519-525.

[23]Sun, Q., Chen, X., Yu, J., et al., 2013. Immune modulatory function of abundant immune-related microRNAs in microvesicles from bovine colostrum. Protein Cell, 4(3):197-210.

[24]Wang, G., Szeto, C.C., 2013. Methods of microRNA quantification in urinary sediment. Methods Mol. Biol., 1024:211-220.

[25]Warensjö, E., Jansson, J.H., Cederholm, T., et al., 2010. Biomarkers of milk fat and the risk of myocardial infarction in men and women: a prospective, matched case-control study. Am. J. Clin. Nutr., 92(1):194-202.

[26]Weber, J.A., Baxter, D.H., Zhang, S., et al., 2010. The microRNA spectrum in 12 body fluids. Clin. Chem., 56(11):1733-1741.

[27]Zhang, L., Hou, D., Chen, X., et al., 2011. Exogenous plant MIR168a specifically targets mammalian LDLRAP1: evidence of cross-kingdom regulation by microRNA. Cell Res., 22(1):107-126.

[28]Zhang, W.Y., Liu, W., Zhou, Y.M., et al., 2012. Altered microRNA expression profile with miR-27b down-regulation correlated with disease activity of oral lichen planus. Oral Dis., 18(3):265-270.

[29]Zhou, Q., Li, M.Z., Wang, X.Y., et al., 2012. Immune-related microRNAs are abundant in breast milk exosomes. Int. J. Biol. Sci., 8(1):118-123.

Open peer comments: Debate/Discuss/Question/Opinion


Please provide your name, email address and a comment

Journal of Zhejiang University-SCIENCE, 38 Zheda Road, Hangzhou 310027, China
Tel: +86-571-87952783; E-mail: cjzhang@zju.edu.cn
Copyright © 2000 - 2024 Journal of Zhejiang University-SCIENCE