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Total RNA were isolated from 200 µL of plasma with the miRNeasy Serum/Plasma Kit from QIAGEN Benelux B.V. (Venlo, Netherlands) following the manufacturer protocol. For technical normalization, 5.6 × 108 copies of a nonhuman synthetic microRNA (miRNeasy Serum/Plasma Spike-In Control, C. elegans miR-39 miRNA mimic) were added to plasma as a spike-in control during the extraction step, before the chloroform addition, as advised by the manufacturer. The final total RNA fraction was eluted in 15 µL of RNase-free water.
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