The Plasma/Serum Circulating and Exosomal RNA Purification Kit (Slurry Format) provides a fast, reliable and simple procedure for isolating circulating RNA and exosomal RNA from plasma and serum samples ranging from 0.25 mL to 5 mL. Free-circulating plasma and serum RNA can serve as both tumor- and fetal-specific markers for cancer detection and prenatal diagnosis. As well, free-circulating RNAs have the potential to provide biomarkers for other disease states. Free-circulating RNA in plasma or serum are usually present as short fragments of less than 1000nt, and free-circulating miRNA (21nt) can also be found in plasma and serum. This kit is able to isolate all sizes of circulating and exosmal RNA, including microRNA, without the use of phenol or chloroform. The slurry format provides an advantage over other available kits in that it does not require extension tubes for the purification of free-circulating and exosomal RNA from large sample volumes. RNA can be isolated from either fresh or frozen samples using this kit. This kit is suitable for the isolation of RNA from serum or plasma prepared from blood collected only on either EDTA or citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications including RT-PCR. Purification is based on using Norgen's proprietary resin as the separation matrix. For the isolation of circulating RNA the plasma/serum samples are mixed with PS Solution A, which contains the separation matrix. PS Solution B and ethanol are then added, and the mixture is centrifuged. PS Solution C is then added to the pellet, and the slurry is loaded onto a provided Mini Filter Spin Columns (BIND). This is followed by washing of the bound RNA to remove the remaining proteins and other impurities (WASH). Lastly, the purified circulating RNA is eluted into the provided Elution Buffer (ELUTE). Please see the procedure flowchart to the right. Preparation time for a single sample is less than 40 minutes. The purified plasma/serum free-circulating and exosmal RNA is eluted in an elution solution that is compatible with real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.
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