EXTRAClean Urine Exosome and Free-Circulating RNA Isolation Kit

Overview

Versatile sample input ranges.
The purified exosomal RNA is free from any circulating RNA-binding proteins.
No phenol extractions, Proteinase K treatment nor carrier RNA required.
No time-consuming ultracentrifugation, filtration nor special syringes are required.
No precipitation reagents nor overnight incubation required.
Concentrate isolated exosomal RNA and free-circulating 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 resin separation matrix.

Norgen’s EXTRAClean Urine 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 urine sample volumes ranging from 250 μL to 30 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 it does not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or 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 various urine volumes using EXTRAClean Urine 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 urine volumes using EXTRAClean Urine 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 urine volumes using EXTRAClean Urine 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 various urine volumes using EXTRAClean Urine 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 urine volumes using EXTRAClean Urine Exosome and 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 urine volumes using EXTRAClean Urine Exosome and Free-Circulating RNA Isolation Midi Kit.

Figure 7. Average read quality distribution relative to raw reads of small RNA sequencing of exosomal RNA extracted from various urine volumes using EXTRAClean Urine 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 urine volumes using EXTRAClean Urine 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 urine volumes using EXTRAClean Urine Exosome and Free-Circulating RNA Isolation Maxi Kit.

Kit Specifications (Spin Column)
Minimum Urine Input	250 μg
Maximum Urine 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 buffers 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.