Extracellular Vesicle RNA Isolation Protocol

pEVs were processed for RNA isolation using the “Maxwell RSC miRNA plasma and serum kit” (Cat# AS1680; Promega). Three synthetic spike-ins (Ath-miR-159a, Cel-miR-254, osa-miR-414) were added to samples after lysis to assess RNA isolation efficiency. Isolated RNA was subjected to quality control for hemolysis detection using the ratio of miR-23a to miR-451 [26 (link)–28 (link)]. Two microliters of RNA were reverse transcribed to complementary DNA (cDNA) using the TaqMan Advanced MicroRNA cDNA synthesis Kit (Cat# A28007; ThermoFisher Scientific, Rockford, USA) according to the manufacturer's protocol.

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Sabato C., Noviello T.M., Covre A., Coral S., Caruso F.P., Besharat Z.M., Splendiani E., Masuelli L., Battistelli C., Vacca A., Catanzaro G., Po A., Anichini A., Maio M., Ceccarelli M., Di Giacomo A.M, & Ferretti E. (2022). A novel microRNA signature for the detection of melanoma by liquid biopsy. Journal of Translational Medicine, 20, 469.

Publication 2022

Maxwell RSC miRNA plasma and serum kit TaqMan Advanced MicroRNA cDNA synthesis Kit

Cdna Hemolysis Microrna Plasma Promega Serum Synthesis

Corresponding Organization :

Other organizations : Sapienza University of Rome, Biogem, University of Siena, Fondazione IRCCS Istituto Nazionale dei Tumori, University of Naples Federico II

Top 5 similar protocols

Variable analysis

independent variables

Ath-miR-159a, Cel-miR-254, osa-miR-414 spike-ins

dependent variables

RNA isolation efficiency
Hemolysis detection using the ratio of miR-23a to miR-451

control variables

RNA isolation using the Maxwell RSC miRNA plasma and serum kit
Reverse transcription of RNA to cDNA using the TaqMan Advanced MicroRNA cDNA synthesis Kit

positive controls

Ath-miR-159a, Cel-miR-254, osa-miR-414 spike-ins

negative controls

Not specified

Annotations

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