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To minimize the effect of RNase on the stability of miRNAs, all glassware and centrifuge tubes were autoclaved, all solutions were treated with diethylpyrocarbon- ate, and all surfaces were decontaminated with RNaseZap (Ambion, TX). RNase-free ultrapure water was used throughout. Reference values of miRNA expression levels in the samples were obtained by using a commercial miRNA qRT- PCR kit from Agilent Technologies (Santa Clara, U.S.). |
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