Quantification of miRNA in Cells, Circulation, and Exosomes

QRT-PCR assay was constructed as previously described [24 (link), 47 ]. Briefly, total RNA in different cells was isolated with TRIzol reagent (Invitrogen), circulating miRNA was extracted using the the miRcute Serum/Plasma miRNA isolation Kit (TIANGEN, DP501), and SEVs miRNAs were isolated by exosomal miRNA and Total Exosome RNA Kit (Ambion) and MirVana RNA isolation kit (Ambion). Primers for miRNAs were designed by Biomics Biotechnologies, whereas primers for macrophage polarization was purchased from Guangzhou RiboBio. Cellular RNA level were normalized to RNU6, while circulating and SEVs miRNAs were normalized to cel-miR-39 (synthetic spike control).

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Zhang C., Chen W., Pan S., Zhang S., Xie H., Zhang Z., Lei W., Bao L, & You Y. (2023). SEVs-mediated miR-6750 transfer inhibits pre-metastatic niche formation in nasopharyngeal carcinoma by targeting M6PR. Cell Death Discovery, 9, 2.

Publication 2023

TRIzol reagent miRcute Serum/Plasma miRNA isolation Kit Total Exosome RNA Kit MirVana RNA isolation kit

Assay Cellular Different cells Exosome Isolation Macrophage polarization Mirnas Plasma Primers Serum Trizol

Corresponding Organization :

Other organizations : Affiliated Hospital of Nantong University, Nantong University

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Variable analysis

independent variables

Different cells

dependent variables

Total RNA level
Circulating miRNA level
SEVs miRNA level

control variables

RNU6 for normalizing cellular RNA level
Cel-miR-39 (synthetic spike control) for normalizing circulating and SEVs miRNAs

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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