Description
The EZBioscience® Fast Tissue RNA Purification Kit provides a simple, reliable, and rapid method for isolating high-quality total RNA from animal tissues and cells, providing high sample processing capability to deal with difficult specimens. Genomic DNA contamination is effectively removed using DNase treatment. The purified total RNA is ready to use and is ideally suitable for downstream applications that are sensitive to low amounts of DNA contamination, such as quantitative, real-time RT-PCR. The purification RNA can also be used in other applications including: RT-PCR, Northern blotting, nuclease protection assays, cDNA library preparation after poly (A)+ selection, and so on. Samples are first lysed and homogenized in a highly denaturing guanidine-isothiocyanate-containing buffer, which immediately inactivates RNases to ensure isolation of intact RNA. After homogenization, ethanol is added to the lysate to provide appropriate binding conditions for RNA, and the sample is then applied to an RNA spin column, where total RNA binds to the membrane. Genomic DNA is removed by DNase treatment and Contaminants are effectively removed by subsequent washing. The purified total RNA is then eluted with Elution Buffer.
Components
Components | EZB-RN5 (100 Preps) |
Lysis Buffer | 65 ml |
Wash Buffer 1* | 13 ml |
Wash Buffer 2* | 13 ml |
Elution Buffer | 10 ml |
gDNA Remover | 220 μl |
Spin Columns for RNA (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 contamination). The Lysis Buffer is suggested to be stored at 4°C when the room temperature is >25°C, or for long-term storage.
Experimental Procedure at a Glance
Features
1.Fast procedure delivering high-quality total RNA in ~20 minutes.
2.high Pure RNA without phenol carryover.
3.High-quality RNA for all downstream applications: RT-PCR, RT-qPCR, Northern blotting, RNA-seq, Poly A+ RNA.
4.Compatibility with a variety of tissue types.
Representative Experimental Results
Figure 1. RNA isolated from Adipose Tissues using EZBioscience® Fast Tissue Purification Kit M: 250bp DNA Ladder(left); The RNA purified using the kit, detected by Nanodrop. The data from Figure 1. indicate that, Fast Tissue RNA Purification Kit can completely replace the TRIzol method in RNA purification.
For detailed information, please check the product manual.