Description
The EZBioscience® EZ-press Serum/Plasma RNA Purification Kit provides a simple, reliable, and rapid method for isolating high-quality total RNA (including mRNA, miRNA, lncRNA, circRNA, et.al) from human and animal serum/plasma. The purified total RNA is suitable for use in a variety of downstream applications, including: RT-PCR, RT-qPCR, Northern blotting, RNA sequencing, and so on. Serum/plasma samples are first lysed and homogenized in a strong denaturant and phenol containing buffer, which immediately inactivates RNases to ensure isolation of intact 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 mixed with Supplemental Reagent and ethanol. The liquid is then passed through the RNA binding Spin Column containing a silica-based membrane to which the RNA binds. Impurities are effectively removed by subsequent washing. The purified total RNA is then eluted with Elution Buffer and may be used in a variety of downstream applications.
Components
Components | EZB-RN2 (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.
Experimental Procedure at a Glance
Features
1. Easy-to-follow protocol for lysis and homogenization.
2. High yields of RNA from human and animal serum/plasma
3. High-quality RNA for all downstream applications: RT-PCR, RT-qPCR, Northern blotting, RNA-seq,, and so on.
Representative Experimental Results
Figure 1. RNA isolated from serum using EZBioscience® EZ-press Serum/Plasma RNA Purification Kit. This qPCR detections results shows that this kit can be well used for the purification of miRNA.
For detailed information, please check the product manual.
