Plasma miRNA Isolation and Quantification

The miRNeasy Mini Kit (QIAGEN, Germany) was applied to isolate total RNA, including small RNAs, from plasma. qPCR analysis was performed as described previously (Mussbacher et al., 2020 (link)). Robustness of RNA extraction, complementary DNA synthesis, and qPCR amplification was assessed using combinations of synthetic spike-in controls (Exiqon, Denmark). Hemolysis was assessed using the ratios of miRNA-23a-3p and miRNA-451a and positive samples were excluded from the analysis.

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Heber S., Laky M., Anscheringer I., Wolschner L., Mussbacher M., Krammer T., Haririan H., Schrottmaier W.C., Volf I., Hackl M., Moritz A, & Assinger A. (2021). Platelet Activation Is Not Always Associated With Platelet-Related Plasma microRNA Abundance – Results From a Randomized Controlled Trial of Periodontal Patients. Frontiers in Physiology, 12, 613515.

Publication 2021

miRNeasy Mini Kit synthetic spike-in controls

Complementary dna Dna synthesis Hemolysis Mirna Plasma Synthesis

Corresponding Organization : Medical University of Vienna

Other organizations : University of Graz

Top 5 similar protocols

Variable analysis

independent variables

Combinations of synthetic spike-in controls (Exiqon, Denmark)

dependent variables

Robustness of RNA extraction
Robustness of complementary DNA synthesis
Robustness of qPCR amplification

control variables

Hemolysis assessed using the ratios of miRNA-23a-3p and miRNA-451a

positive controls

Synthetic spike-in controls (Exiqon, Denmark)

negative controls

Not explicitly mentioned

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