miRNAs were isolated from serum by the miRNeasy Serum/Plasma Advanced Kit and from ECs by MasterPure™ Complete DNA & RNA Purification Kit (Lucigen), retrotranscribed using the miRCURY LNA RT Kit, and quantified by qPCR with the miRCURY LNA SYBR Green PCR Kit (all from Qiagen) as their protocols indicate. Specific primers (Qiagen) were used to measure miRNAs of interest, and the 2‐ΔΔCT method was applied to quantitate the different data15 (link) obtained in the LightCycler 96 Real‐Time PCR System (Roche Life Science).
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Quantitative miRNA Analysis from Serum and Cells
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Corresponding Organization : Universidad Alfonso X el Sabio
Other organizations : Hospital Universitario Fundación Jiménez Díaz, Autonomous University of Madrid, Hospital Infantil Universitario Niño Jesús, Universidad Complutense de Madrid, Hospital Universitario Puerta de Hierro Majadahonda, Hospital Central de la Cruz Roja San José y Santa Adela, Centre for Plant Biotechnology and Genomics, Universidad Politécnica de Madrid, City University of New York, The Graduate Center, CUNY, Lehman College
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Variable analysis
independent variables
- Isolation of miRNAs from serum by the miRNeasy Serum/Plasma Advanced Kit
- Isolation of miRNAs from ECs by MasterPure™ Complete DNA & RNA Purification Kit (Lucigen)
- Retrotranscription of miRNAs using the miRCURY LNA RT Kit
- Quantification of miRNAs by qPCR with the miRCURY LNA SYBR Green PCR Kit (Qiagen)
- Use of specific primers (Qiagen) to measure miRNAs of interest
- Levels of miRNAs of interest
- Protocols provided by the manufacturers (Qiagen, Lucigen) for the respective kits and reagents used
- Not explicitly mentioned
- Not explicitly mentioned
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