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. 2016 May 3;9:31. doi: 10.3389/fnmol.2016.00031

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Copyright © 2016 Piscopo, Grasso, Fontana, Crestini, Puopolo, Del Vescovo, Venerosi, Calamandrei, Vencken, Greene, Confaloni and Denti.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution and reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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mRNA:miRNA isolation technique for GRN mRNA from SK-N-BE cells. SK-N-BE 3′UTR sequencing showing a TC genotype (A). A capture anti-sense DNA oligonucleotide with a biotin modification at the 5′ end was designed to pull-down GRN mRNA. RT-qPCR showed enrichment of GRN mRNA and miR-659-3p in SK-N-BE (B,C, respectively) and Kelly samples (D,E, respectively) compared to a scramble oligonucleotide used as negative control. mRNA and miRNA expression was quantified using the 2^−ΔCt method. Mean ± SEM of three biological replicates is shown (N = 9, *p < 0.05; **p < 0.01; ***p < 0.001 vs. scramble non specific control).