Urine Exosome RNA Isolation Kit

(Cat# 47200)

A rapid procedure for the isolation of exosomal RNA from urine sample

Isolation of exosomal RNA molecules from urine samples
Rapid and convenient spin-column protocol
Isolate inhibitor-free urinary microRNA

Product Description
Product Description
Applications & Data
Applications & Data
Specifications
Specifications
Resources
Resources
Ordering
Ordering

Norgen’s Urine Exosome RNA Isolation Kit constitutes an all-in-one system for the concentration and isolation of exosomal RNA from urine and tissue culture media. Purification is based on spin column chromatography using Norgen’s proprietary resin as the separation matrix. Briefly, the urine samples are first centrifuged in order to remove all cells. The urine exosomes are then separated from the urine using Norgen's proprietary resin. Next, the exosomes are lysed to release the RNA, which is then bound to Norgen's resin (BIND). Under these conditions only the RNA will bind to Norgen's resin while the remaining contaminants are removed in the flowthrough or retained on top of the resin. The bound RNA is then washed to remove any remaining impurities (WASH). Lastly, the purified exosome RNA is eluted into 100 µL of the provided Elution Buffer (ELUTE). Please see procedure flowchart to the right.

The exosomal RNA is isolated free from inhibitors and can be used as template in a number of sensitive downstream assays. The kit is designed to allow for the isolation of exosomal RNA from 50 urine samples. The kit is designed to allow for the isolation of exosomal RNA from 1 mL to 10 mL urine samples.

FEATURES AND BENEFITS

Fast and easy processing - Rapid spin-column format allows for the processing of multiple samples in under 30 minutes.
Versatile urine input volume - Isolate urine exosomal RNA from 1 mL to 10 mL of urine samples.
No phenol:chloroform extractions - Norgen’s Urine Exosome RNA Isolation Kit isolates RNA without the use of harmful chemicals such as phenol or chloroform.
Isolate inhibitor-free RNA suitable for downstream applications - Purified RNA is fully compatible with various downstream applications relating to gene regulation and functional analysis, including RT-PCR, northern blotting and microarray analysis.

APPLICATIONS

The purified urine exosome RNA is compatible with most of the molecular biological applications such as:

Bioanalyzer
Quantitative, real-time RT-PCR for
RT-PCR
Northern blotting
RNase protection
Primer extension
Expression array assays
Next Generation Sequencing
RNA Cloning

Figure 1. Isolation of Urine Exosomal RNA from Different Urine Volumes. Norgen’s Urine Exosome RNA Isolation Kit was used to isolate urine exosomal RNA from different urine volumes ranging from 1 – 15 mL, and the yield of RNA obtained was compared to the conventional Nanofilteration method. Norgen’s Urine Exosome RNA Isolation Kit procedure showed a linear increase in the RNA yield with an increase in urine volume processed. Furthermore, when isolating exosomal RNA from 10 mL of urine using Norgen's kit and the conventional Nanofilteration method, Norgen's Urine Exosome RNA Isolation Kit showed a higher yield.

Figure 2. Detection of Urine Exosomal RNA Isolated from Different Urine Volumes. Norgen’s Urine Exosome RNA Isolation Kit was then used to isolate Urine Exosomal RNA from different urine volumes rangeing from 1 – 15mL Urine. The purified urine exosomal RNA was then used as the template in an RT-qPCR reaction to detect the human 5S gene. Three microlitres of the isolated urine exosomal RNA was used as the template in the RT step, and 3 μL from the RT step was used in the qPCR reaction. As it can be seen, the qPCR was able to successfully detect and amplify the 5S gene from RNA isolated from different urine volumes, indicating the high quality of the isolated urine exosomal RNA.

Figure 3: Effective and Consistent Detection of Urine Exosome RNA. Norgen's Urine Exosome RNA Isolation Kit can effectively isolate RNA from urine. Urine Exosome RNA was isolated from 5 mL of human cell-free urine using Norgen's Urine Exosome RNA Isolation Kit (blue), a ExoQuick-TC Exosome Precipitation Reagent (green) and a Amicon® Ultra-15 Filter (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 Urine Exosome RNA Isolation Kit is the only product that showed consistent detection of all tested transcripts with the highest quality.

PROTOCOL

Urine Exosome RNA Isolation Kit - Protocol
PDF version

PRODUCT INFORMATION SHEETS

Urine Exosome RNA Isolation Kit - Product Information Sheet
PDF version

APPLICATION NOTES

POSTERS

Comparative Study on the Isolation of Exosomal RNA From Urine Using Different Methods
PDF version

MSDS

SUPPLEMENTARY PROTOCOLS

REFERENCES

FREQUENTLY ASKED QUESTIONS