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Urine Exosome Total RNA Isolation Kit Add to Cart

Cat#: 47200-NB
Quantity: 50 preps
Price: 503 €
Supplier: Norgen
Shipping: RT
User Manual  

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

• 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 or chloroform extractions
• Isolate inhibitor-free RNA suitable for downstream applications

Norgen’s Urine Exosome RNA Isolation Kit provides a rapid procedure for the isolation of exosomal RNA from urine samples. Exosomes are 40 - 100 nm membrane vesicles, which are secreted by most cell types. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascite fluids, among other biological fluids. Evidence has been accumulating recently that these vesicles act as cellular messengers, conveying information to distant cells and tissues within the body. The exosomes contain cell-specific proteins, lipids and RNAs, which are transported to other cells, where they can alter function and/or physiology. These exosomes may play a functional role in mediating adaptive immune responses to infectious agents and tumours, tissue repair, neural communication and transfer of pathogenic proteins. Recent work has demonstrated the presence of distinct subsets of microRNAs within exosomes which depend upon the tumour cell type from which they are secreted. For this reason exosomal RNAs may serve as biomarkers for various diseases including cancer. As the RNA molecules encapsulated within exosomes are protected from degradation by RNAses they can be efficiently recovered from biological fluids, such as urine. Norgen’s Urine Exosome RNA Isolation Kit makes exosomal RNA discoveries simple, rapid and reliable. Users can simultaneously concentrate and isolate high quality exosomal RNA, including microRNA, fully compatible with various downstream applications relating to gene regulation and functional analysis, including RT-PCR, northern blotting and microarray analysis.

Workflow
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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.


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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 ìL 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.


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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.


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