Description
The EZBioscience® EZ-press microRNA Purification Kit PLUS provides a simple, reliable, and rapid method for isolating high-quality RNA (total RNA including miRNA) from a wide variety of sources, without the need for toxic substances such as phenol or chloroform. This kit can be used with cultured cells and part of animal tissues. Biological samples are first lysed and homogenized in a strong denaturant containing buffer, which immediately inactivates RNases to ensure isolation of intact RNA. After homogenization, the lysate is passed through the DNA Removing Spin Column which traps the DNA on the column, while RNA can pass through the column with the flow-through. Then ethanol is added to the flow-through. The mixture is then passed through the RNA binding Spin Column to which the RNA binds. Impurities are effectively removed by subsequent DNase treatment and washing. 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, and so on.
Components
Components | B0005-plus (50 Preps) |
Lysis Buffer | 30 ml |
Wash Buffer* | 8 ml |
Elution Buffer | 25 ml |
gDNA Remover | 110 μl |
DNA Removing Spin Columns (with Collection Tubes) | 50 Preps |
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.
Storage
Store the gDNA Remover at -20°C. Store other components at room temperature (When using these buffers, be careful to avoid of contamination).
Experimental Procedure at a Glance
Features
1. Fast and simple procedure delivering high-quality RNA (total RNA containing miRNA) in ~15 minutes.
2. Efficient gDNA removal with unique DNA Removing Spin Columns and gDNA Remover.
3. Safe and non-toxic: no phenol/chloroform extraction, or ethanol precipitation.
4. High-quality RNA for downstream applications: RT-PCR, RT-qPCR, RNA Sequencing, and so on.
Representative Experimental Results
Figure 1. Detection of microRNA expression in A549 cells by RT-qPCR (the microRNA was purified by EZBioscience® EZ-press microRNA Purification Kit PLUS). These results showed that both the miR-145 and U6 can be amplified efficiently and specially.
For detailed information, please check the product manual.
