Plasma/Serum Circulating and Exosomal RNA Purification 96-well Kit (Slurry Format), 0.25 - 2 ml sample input

Product Description

• Isolate all sizes of circulating RNA
• Process 96 samples in less than 1 hour
• Concentrate circulating RNA into small elution volumes
• Isolate inhibitor-free circulating RNA

Norgen’s Plasma/Serum Circulating RNA Purification 96-Well Kit (Slurry Format) provides a fast, reliable and simple procedure for isolating circulating RNA from plasma and serum samples ranging from 0.25 ml to 2 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 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 RNA from large sample volumes. RNA can be isolated from either fresh or frozen samples using this kit. The kit allows for the concentration of RNA that is present in low concentrations (1-100ng/ml circulating RNA in human plasma).

Purification is based on using Norgen's proprietary resin as the separation matrix. For the isolation of circulating RNA the plasma/serum samples are first mixed with PS Solution A, which contains the separation matrix, and PS Solution B. Next, ethanol and PS Solution C are added and the slurry is loaded onto a provided 96-Well Filter Plate. This is followed by washing of the bound RNA to remove the remaining proteins and other impurities. Lastly, the purified circulating RNA is eluted into 100 µl of the provided Elution Buffer. The procedure can be performed using either a vacuum manifold or centrifugation. Please see the procedure flowchart to the right.

Preparation time for 96 samples is approximately 1 hour. The purified plasma/serum free-circulating 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.

Performance Data

Figure 2. Effective Isolation of Plasma Circulating RNA from Different Volumes. Norgen's Plasma/Serum Circulating RNA Purification Mini Kit (Slurry Format) was used to isolate circulating RNA from 0.5mL, 1mL and 2mL plasma. Three microlitres of the purified RNA was then used as the template in RT-qPCR reactions to detect the human 5S gene. The 5S housekeeping gene was detected from all plasma sample volumes used. The amplification of the 5S rRNA showed an increasing amount of RNA with increasing the sample input volume. This is represented by the decrease of the Ct value with increasing the sample input volume. Average Ct. values for the amplification of the 5S rRNA isolated from 0.5mL plasma is represented by the Red bar, Average Ct. values for the amplification of the 5S rRNA isolated from 1mL plasma is represented by the Green bar whereas Average Ct. values for the amplification of the 5S rRNA isolated from 2mL plasma is represented by the blue bar.