EXTRAClean Plasma/Serum Exosome and Free-Circulating RNA Isolation Kit

Overview

The purified exosomal RNA is free from any circulating RNA-binding proteins.
No phenol extractions, Proteinase K treatment, nor carrier RNA.
No time-consuming ultracentrifugation, filtration nor special syringes are required.
Concentrate isolated RNA into a flexible elution volume ranging from 50 μL to 100 μL.
Purification is based on EXTRAClean spin column chromatography that uses Norgen’s proprietary separation matrix.

Norgen’s EXTRAClean Plasma/Serum Exosome and Free-Circulating RNA Isolation Kit constitutes an all-in-one system for the purification of exosomes and the sequential isolation of RNA and free-circulating RNA from different plasma/serum sample volumes ranging from 50 μL and up to 10 mL. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. The EXTRAClean columns undergo stringent processing and rigorous quality control measures to minimize contamination traces, ensuring optimal results for sensitive applications such as NGS. The kit is designed to isolate all sizes of RNA, including microRNA. The kit provides a clear advantage over other available kits in that they do not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or any protease treatments. Moreover, the kit allows the user to elute into a flexible elution volume ranging from 50 μL to 100 μL. The RNA isolated from the purified exosomes is free from any protein-bound circulating RNA and is of the highest integrity. Moreover, the free-circulating, protein-bound, RNA is free from any exosomal RNA. The purified RNA can be used in a number of downstream applications including real time PCR, NGS application, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.

Details

Supporting Data

Figure 1. Average read quality distribution relative to raw reads of small RNA sequencing of exosomal RNA extracted from plasma and serum samples using EXTRAClean Plasma/Serum Exosome and Free-Circulating RNA Isolation Mini Kit. Calculations were relative to raw reads.

Figure 2. Genome mapping distribution relative to input reads obtained from small RNA sequencing of exosomal RNA extracted from various plasma and serum samples using EXTRAClean Plasma/Serum Exosome and Free-Circulating RNA Isolation Mini Kit.

Figure 3. Average small RNA biotype distribution relative to the input reads obtained from sequencing of exosomal RNA extracted from various plasma and serum using EXTRAClean Plasma/Serum Exosome and Free-Circulating RNA Isolation Mini Kit.

Figure 4. Average read quality distribution relative to raw reads of small RNA sequencing of exosomal RNA extracted from plasma and serum samples using EXTRAClean Plasma/Serum Exosome and Free-Circulating RNA Isolation Midi Kit. Calculations were relative to raw reads.

Figure 5. Genome mapping distribution relative to input reads obtained from small RNA sequencing of exosomal RNA extracted from various plasma and serum samples using EXTRAClean Plasma/Serum Exosomeand Free-Circulating RNA Isolation Midi Kit.

Figure 6. Average small RNA biotype distribution relative to the input reads obtained from sequencing of exosomal RNA extracted from various plasma and serum using EXTRAClean Plasma/Serum Exosome and Free-Circulating RNA Isolation Maxi Kit.

Figure 7. Average read quality distribution relative to raw reads of small RNA sequencing of exosomal RNA extracted from plasma and serum samples using EXTRAClean Plasma/Serum Exosome and Free-Circulating RNA Isolation Maxi Kit. Calculations were relative to raw reads.

Figure 8. Genome mapping distribution relative to input reads obtained from small RNA sequencing of exosomal RNA extracted from various plasma and serum samples using EXTRAClean Plasma/Serum Exosome and Free-Circulating RNA Isolation Maxi Kit.

Figure 9. Average small RNA biotype distribution relative to the input reads obtained from sequencing of exosomal RNA extracted from various plasma and serum using EXTRAClean Plasma/Serum Exosome and Free-Circulating RNA Isolation Maxi Kit.

Kit Specifications (Spin Column)
Minimum Plasma Input	50 uL
Maximum Plasma Input	1 mL
Size of RNA Purified	All sizes, including miRNA and small RNA (< 200 nt)
Elution Volume	50-100 μL
Time to Complete 10 Purifications	35-40 minutes
Average Yields	Variable depending on specimen

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.