Description
The EZBioscience® Universal microRNA Purification Kit integrates phenol/guanidine-based sample lysis and silica-membrane purification of high-quality RNA from animal tissues and cells. Samples are first lysed and homogenized in Lysis 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 ethanol is added to provide appropriate binding conditions. The sample is then applied to the RNA Spin Column, where the 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, RNA Sequencing, RNA chip and so on.
Components
Components | EZB-miRN1 (50 Preps) |
Lysis Buffer | 30 ml |
Buffer A | 6 ml |
Wash Buffer 1* | 8 ml |
Wash Buffer 2* | 8 ml |
Elution Buffer | 25 ml |
Spin Columns (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 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 (total RNA containing miRNA) from animal tissues and cells.
3. High-quality RNA for many downstream applications: RT-PCR, RT-qPCR, Northern blotting, RNA-seq, and so on.
Representative Experimental Results
Figure 1. microRNA is isolated using EZBioscience® Universal microRNA Purification Kit. As shown in the following figure, the amplification curve, melt curve and Ct value of these microRNAs were very good, indicating that the expression of microRNA can be well detected using this kit.
For detailed information, please check the product manual.
