Description
The EZBioscience® EZ-press RNA Purification Kit provides a simple, reliable, and rapid method for isolating high-quality total RNA from a wide variety of sources, without the need for toxic substances such as phenol or chloroform. The Kit can be used with cultured cells. 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, ethanol is added to the lysate, creating conditions that promote selective binding of RNA to the silica-based membrane. The sample is then applied to the Spin Column, where the total RNA binds to the membrane, then the membrane is treated with gDNA Remover to eliminate the DNA. Contaminants are efficiently washed away by subsequent step of washing. The high-quality total RNA is then eluted in Elution Buffer. The purified total RNA is suitable for use in a variety of downstream applications, including: RT-PCR, RT-qPCR, and so on.
Components
Components | B0004DP (100 Preps) |
Lysis Buffer | 55 ml |
Wash Buffer* | 13 ml |
Elution Buffer | 10 ml |
gDNA Remover | 220 μl |
Spin Columns (with Collection Tubes) | 100 Preps |
*Before using for the first time, add 52 ml 100% ethanol to the Wash Buffer and mix thoroughly.
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 procedure delivering high-quality total RNA in ~8 minutes.
2. Safe and non-toxic: no phenol/chloroform extraction, or ethanol precipitation.
3. High-quality RNA for downstream applications: RT-PCR, RT-qPCR, and so on.
Representative experimental results
Figure 1. RNA isolated from different amounts of A549 cells using EZBioscience® EZ-press RNA Purification Kit and TRIzol. M: 1kb DNA Ladder; Lanes 1 ~ 3: EZ-press RNA Purification Kit; Lanes 4 ~ 6: TRIzol; 1: 2 × 105 cells; 2: 4 × 105 cells; 3: 6 × 105 cells; 4: 2 × 105 cells; 5: 4 × 105 cells; 6: 6 × 105 cells. These results showed that more RNA can be isolated from the same amount of cells by EZBioscience® EZ-press RNA Purification Kit than TRIzol.
Figure 2. Detection of mRNA level of 4 genes (ACTIN, P53, SNAIL and ZO-1) in A549 cells by RT-qPCR (the RNA was purified by EZBioscience® EZ-press RNA Purification Kit and TRIzol method respectively). Figure 2. Shows that, the Ct values of these genes in the RNA sample purified by EZ-press RNA Purification Kit are highly correlated with the RNA purified by TRIzol method (R2=0.9898). And the Ct values from the RNA derived by EZ-press RNA Purification Kit are smaller than by the TRIzol method.
These data from Figure 1. and Figure 2. indicate that, EZBioscience® EZ-press RNA Purification Kit can be a good substitution for the TRIzol method in RNA purification. And also, using this Kit can get better results than the TRIzol method.
For detailed information, please check the product manual.
