Description
The EZBioscience® Universal RNA Purification Kit integrates phenol/guanidine-based sample lysis and silica-membrane purification of high-quality total 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 total RNA binds to the membrane, and contaminants are effectively washed away. Finally, high-quality RNA is eluted in Elution Buffer. The purified total RNA is suitable for use in a variety of downstream applications, including: RT-PCR, RT-qPCR, RNA-seq, Northern blotting, Poly A+ RNA selection, Microarray analysis, and so on.
Components
Components | EZB-RN4 (100 Preps) |
Lysis Buffer | 55 ml |
Buffer A | 12 ml |
Wash Buffer 1* | 13 ml |
Wash Buffer 2* | 13 ml |
Elution Buffer | 25 ml |
Spin Columns (with Collection Tubes) | 100 Preps |
*Before using for the first time, add 52 ml of 100% ethanol to the Wash Buffer 1, 52 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. The kit contains Buffer A, which can replace chloroform.
3. High yields of RNA from animal tissues and cultured cells.
4. High-quality RNA for all downstream applications: RT-PCR, RT-qPCR, Northern blotting, RNA-seq, Poly A+ RNA, and so on.
Representative Experimental Results
Figure 1. RNA isolated from 30 mg mouse muscle using EZBioscience® Universal RNA Purification Kit and TRIzol (eluted in 30 μl of Elution Buffer each, 5 μl RNA of each is loaded for electrophoresis). M: 250bp DNA Ladder; Lanes 1: TRIzol; Lanes 2: EZBioscience® Universal RNA Purification Kit; These results showed that more RNA can be isolated from the same amount of tissue by EZBioscience® Universal RNA Purification Kit than TRIzol.
Figure 2. lncRNA isolated from 293T cells are detected. These results showed that, this EZBioscience® Universal RNA Purification Kit can be used for lncRNA purification and achieved good performance.
For detailed information, please check the product manual.
