Introduction
The EZBioscience® Exosome RNA Purification Kit provides a simple, reliable, and rapid method for isolating high-quality total RNA (including miRNA, mRNA, lncRNA and circRNA, etc.) from exosomes, providing high sample processing capability.
Exosomes can be derived from ultracentrifugation or isolated by exosome isolation kit, such as EZBioscience® Exosome Isolation Kit (from plasma/serum, Cat. No.: EZB-exo1) /Exosome Isolation Kit (from cell culture medium, Cat. No.: EZB-exo2).
Exosome samples are first lysed in a strong denaturant and phenol containing buffer, which immediately inactivates RNases to ensure isolation of intact total RNA. After addition of Buffer A, the homogenate is separated into aqueous and organic phases by centrifugation. RNA partitions to the upper, aqueous phase while DNA partitions to the interphase and proteins to the lower, organic phase or the interphase. The upper, aqueous phase is collected, and ethanol is added to provide appropriate binding conditions. The sample is then applied to the RNA Spin Column, where the total RNA binds to the membrane, and contaminants are effectively washed away. Finally, high-quality RNA is eluted in Elution Buffer. The purified RNA is suitable for use in a variety of downstream applications, including: RT-PCR, RT-qPCR, Northern blotting, RNA sequencing, RNA Chip, and so on.
Components
Component | EZB-exo-RN1 (50 Preps) |
Lysis Buffer | 30 ml |
| Buffer A | 12 ml |
| Supplemental Reagent | 550 μl |
Wash Buffer 1* | 8 ml |
Wash Buffer 2* | 8 ml |
Elution Buffer | 10 ml |
Spin Columns for RNA (with Collection Tubes) | 50 Preps |
*Before using for the first time, add 32 ml of 100% ethanol to the Wash Buffer 1, 32 ml of 100% ethanol to the Wash Buffer 2.
Storage
Store the Lysis Buffer and Buffer A at 2 ~ 8°C, protect from light. Store the Supplemental Reagent at -20°C. Store other components at room temperature (When using these buffers, be careful to avoid of contamination). Divide the Elution Buffer into small aliquots upon reception is suggested.
Note:
The concentrations of the most exosome RNAs are always too low to be measured by common microscale ultraviolet spectrophotometer, such as Nanodrop. If you need to measure the exosome RNA concentration, the Qubit (from Thermo Fisher) is suggested. In most experiments, the measurement of the exosome RNA concentration is unnecessary, such as qRT-PCR. You just need to make sure to use exosomes isolated from equal samples.
For detailed information, please check the product manual.
