Overview

Norgen’s Plasma/Serum Circulating and Exosomal RNA Purification Kit Dx (Slurry Format) provides a fast, reliable and simple procedure for isolating circulating and exosomal RNA from plasma/serum samples for subsequent in vitro diagnostic use. Purification is based on the use of Norgen’s proprietary resin as the separation matrix, and the kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to small RNAs.

This kit is designed to be used with any downstream application employing enzymatic amplification or other enzymatic modifications of RNA followed by signal detection or amplification. Any diagnostic results generated using the RNA isolated with Plasma/Serum Circulating and Exosomal RNA Purification Kit Dx (Slurry Format) in conjunction with an in vitro diagnostic assay should be interpreted with regard to other clinical or laboratory findings.

To minimize irregularities in diagnostic results, suitable controls for downstream applications should be used.

Norgen’s Plasma/Serum Circulating and Exosomal RNA Purification Kit Dx (Slurry Format) is intended for use by professional users such as technicians, physicians and biologists experienced and trained in molecular biological techniques including RNA isolation.

Norgen’s Plasma/Serum Circulating and Exosomal RNA Purification Kit Dx (Slurry Format) does not provide a diagnostic result. It is the sole responsibility of the user to use and validate the kit in conjunction with a downstream in vitro diagnostic assay.

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Figure 1. Isolation and Detection of Circulating RNA from 1 mL Samples of Serum and Plasma Norgen's Plasma/Serum Circulating and Exosomal RNA Purification Kit Dx (Slurry Format) was used to isolate circulating RNA from 1 mL samples of serum and plasma. Five microlitres of the purified RNA was then used as the template in RT-qPCR reactions to detect the human 5S gene and the miR-21 gene. Both the 5S housekeeping gene and the miR-21 gene could be detected from plasma (red) and serum (green), indicating the high quality of the purified RNA. The black line corresponds to the no template control.

Figure 2. Effective and Consistent Detection of Plasma Exosome RNA. Norgen's Plasma/Serum Circulating and Exosomal RNA Isolation Kits can effectively isolate RNA from plasma. Plasma Exosome RNA was isolated from 500 µL of human plasma prepared from blood collected on citrate, EDTA or Heparin in triplicates using Norgen's Plasma/Serum Circulating and Exosomal RNA Isolation Kit Dx (blue), a ExoQuick Exosome Precipitation Reagent (green) and Qiagen’s QIAzol extraction followed by a modified Qiagen’s RNeasy Mini Kit cleanup (red). Stem loop RT-qPCR using primers specific to miR-21 and miR-16 as well as the housekeeping 5S rRNA was performed. In brief, three microliters of the 100 µL isolated RNA was then subjected to a 20 µL reverse transcription using 5S rRNA, miR-21 and miR-16 stem-loop reverse primer or reveres primer. Three microliters of the reverse transcription was used in a 20 µL real-time PCR reaction with primers to detect the human miR-21, human miR-16 and the 5S rRNA. Norgen's Plasma/Serum Circulating and Exosomal RNA Isolation Kit Dx is the only product that showed consistent detection of all tested transcripts with the highest quality regardless the type of the anti-coagulant used for blood collection.

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