Name: Exosomal RNA Isolation Kit
Brand: Norgen Biotek Corp.
SKU: 58000
Price: 405.000000 USD
Availability: InStock

Features and Benefits

Isolate all sizes of exosomal and extracellular vesicle RNA, including microRNA
Bind and elute all RNA irrespective of size or GC content, without bias
No phenol extractions
No Proteinase K treatment
No carrier RNA
Concentrate isolated RNA into a flexible elution volume ranging from 50 µL to 100 µL
Purify high-quality RNA in 15-20 minutes
Purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen's NGS services
Purification is based on spin column chromatography that uses Norgen’s proprietary resin

This kit provides a fast, reliable and convenient method to isolate and concentrate exosomal RNA from previously purified exosomes. It allows for the isolation of RNA from intact extracellular vesicles (EVs) purified from different urine or plasma/serum sample volumes. The purification is based on Norgen’s proprietary resin.

The Exosomal RNA Isolation Kit is designed to isolate all sizes of RNA, including microRNA. Moreover, the kit allows the user to elute into a flexible elution volume ranging from 50 µL to 100 µL. The purified RNA is of the highest integrity, and can be used in a number of downstream applications including real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.

Details

Supporting Data

Figure 1. Isolation of RNA from exosomes purified from different urine volumes. Norgen's Exosomal RNA Isolation Kit (Cat# 58000) was used to isolate RNA from exosomes isolated from different urine volumes purified using Norgen's Urine Exosome Purification Kits (Cat# 57700, 57800 and 57900). 2 μL of the isolated RNA was then used as the template in RT-qPCR reactions to assess the amplification of the isolated urinary exosomal miR-30a. (A) The average Ct value for urinary exosomal miR-30a is linearly decreasing with increasing the sample input volume. B) The relative amount of the urinary exosomal miR-30a shows excellent linearity with a percentage of recovery of more than 90%.

Figure 2. Isolation of RNA from exosomes purified from different plasma volumes. Norgen's Exosomal RNA Isolation Kit (Cat# 58000) was used to isolate RNA from exosomes isolated from different plasma volumes purified using Norgen's Plasma/Serum Exosome Purification Kits (Cat# 57400, 57500 and 57600). 2 μL of the isolated RNA was then used as the template in RT-qPCR reactions to assess the amplification of the isolated plasma exosomal miR-26a. (A) The average Ct value for plasma exosomal miR-26a is linearly decreasing with increasing the sample input volume. B) The relative amount of the plasma exosomal miR-26a shows excellent linearity with a percentage of recovery of more than 90%.

Kit Specifications
Sample type	Exosomes purified using Norgen’s Purification Kits and most other methods
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

*Please check page 5 of the product insert for the average yields and the common RNA quantification methods.

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.

Component	Cat. 58000 (50 preps)
Lysis Buffer A	20 mL
Lysis Additive B	2 mL
Wash Solution A	18 mL
Elution Solution A	6 mL
Mini Spin Columns	50
Collection Tubes	50
Elution Tubes (1.7 mL)	50
Product Insert	1

Documentation

RELATED BLOGS

FAQs

Spin Column

What If a variable speed centrifuge is not available?

A fixed speed centrifuge can be used, however reduced yields may be observed.

At what temperature should I centrifuge my samples?

All centrifugation steps are performed at room temperature. Centrifugation at 4°C will not adversely affect kit performance.

What if I added more or less of the specified reagents’ volume?

Adding more or less than the specified volumes may reduce both the quality and the quantity of the purified RNA. Eluting your RNA in high volumes will increase the yield but will lower the concentration. Eluting in small volumes will increase the concentration but will lower the overall yield.

What if I forgot to do a dry spin before my final elution step?

Your purified RNA will be contaminated with the Wash Solution A. This may reduce the quality of your purified RNA and will interfere with your downstream applications.

Can I perform a second elution?

Yes, but it is recommended that the 2nd elution be in a smaller volume (50% of 1st Elution). It is also recommended to perform the 2nd elution into a separate elution tube to avoid diluting the 1st elution.

Why do my samples show low RNA yield?

Exosomes contain very little RNA. This varies from individual to individual based on numerous variables. In order to increase the yield, the amount of urine, plasma or serum input could be increased.

Why do the A260/280 ratio of the purified RNA is lower than 2.0?

Most of the Exosomal RNA is short RNA fragments with a very low concentration, where the A260/280 ratio tends to decrease with the decrease in the RNA concentration. The A260/280 ratio is normally between 1-1.6. This low A260/280 ratio will not affect any downstream application.

Why does my isolated RNA not perform well in downstream applications?

If a different Elution Buffer was used other than the one provided in the kit, the buffer should be checked for any components that may interfere with the application. Common components that are known to interfere are high salts (including EDTA), detergents and other denaturants. Check the compatibility of your elution buffer with the intended use.

Can I use this kit with exosomes purified with a method other than Norgen's exosome isolation kits?

Yes, this kit can be used with exosomes purified using most other methods as well. The protocol provides instructions to process those samples easily.

Citations

Title	Lipotoxicity-polarised macrophage-derived exosomes regulate mitochondrial fitness through Miro1-mediated mitophagy inhibition and contribute to type 2 diabetes development in mice
Citation	Diabetologia 2023.
Authors	Jian-Ming Li, Xianyu Li, Lawrence W. C. Chan, Ruinian Hu, Tian Zheng, Haojie Li and Sijun Yang

Title	Extracellular Vesicles as Potential Bladder Cancer Biomarkers: Take It or Leave It?
Citation	International journal of molecular science 2023.
Authors	Ana Teixeira-Marques ,Catarina Lourenço ,Miguel Carlos Oliveira ,Rui Henrique and Carmen Jerónimo

Title	Extracellular vesicle-microRNAs mediated response of bovine ovaries to seasonal environmental changes
Citation	Journal of Ovarian Research volume 2023.
Authors	Ahmed Gad, Kamryn Joyce, Nico Graham Menjivar, Daniella Heredia, Camila Santos Rojas, Dawit Tesfaye & Angela Gonella-Diaza

Title	Exosomal miR-let-7c-5p is involved in the cognitive function of type 2 diabetes mellitus patients by interleukin 10: A cross-sectional study
Citation	Journal of diabetes 2023.
Authors	Hui Zhang, Shufang Yang, Wenwen Zhu, Tong Niu, Jue Wang, Mingyue Yang, Enlin Liu, Jumei Wang, Sumei Li, Haoqiang Zhang

Title	Exosomal circTGFBR2 promotes hepatocellular carcinoma progression via enhancing ATG5 mediated protective autophagy.
Citation	cell death & disease 2023.
Authors	Xin Wang, Fenglin Dong, Yingqiao Wang, Honglong Wei, Tao Li, Jie Li

Title	Effect of Storage Conditions on Small Noncoding RNA Concentrations in Human Milk using the Size Exclusion Chromatography Method for Extracellular Vesicle Isolation.
Citation	NHSJS 2023.
Authors	Samantha Wu, Amy L. Non

Title	Dynamics of extracellular vesicle-coupled microRNAs in equine follicular fluid associated with follicle selection and ovulation
Citation	Molecular Human Reproduction, 2023.
Authors	Samuel Gebremedhn, Ahmed Gad, Ghassan M Ishak, Nico G Menjivar, Melba O Gastal, Jean M Feugang, Radek Prochazka, Dawit Tesfaye, Eduardo L Gastal

Title	Differential expression of exosomal miRNAs and proteins in the plasma of systemic lupus erythematous patients
Citation	Heliyon 2023.
Authors	Wencong Song, Chunhong Li, Jie Qiu, Jiyou Dong, Dongzhou Liu, Yong Dai

Title	Circulating Cell-Free RNA in Blood as a Host Response Biomarker for the Detection of Tuberculosis
Citation	Nature Communications 2023.
Authors	Adrienne Chang,Conor J. Loy,Joan S. Lenz, Amy Steadman, Alfred Andama, Nguyen Viet Nhung,Charles Yu, William Worodria, Claudia M. Denkinger, Payam Nahid, Adithya Cattamanchi, and Iwijn De Vlaminck

Title	Extracellular microRNA signature in chronic kidney disease
Citation	American Journal of Physiology - Renal Physiology 2017.
Authors	Muralidharan, J., Ramezani, A., Hubal, M. J., Knoblach, S., Shrivastav, S., Karandish, S., ... & Kopp, J. B.

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Exosomal RNA Isolation Kit