The EZBioscience® EZ-press Tissue RNA Purification Kit provides a simple, reliable, and rapid method for isolating high-quality total RNA from animal tissues, providing high sample processing capability to deal with difficult specimens.
The purified total RNA is suitable for use in a variety of downstream applications, including: RT-PCR, RT-qPCR, Northern blotting, nuclease protection assays, cDNA library preparation after poly (A)+ selection, and so on.
Tissue samples are first lysed and homogenized in a strong denaturant and phenol containing buffer, which immediately inactivates RNases to ensure isolation of intact RNA. After homogenization, ethanol is added to the sample. The lysate is then passed through the RNA binding Spin Column containing a silica-based membrane to which the RNA binds. Impurities are effectively removed by subsequent washing.
The purified total RNA is then eluted in Elution Buffer and may be used in a variety of downstream applications.
Experimental Procedure at a Glance
An overview of the experiment process
Note: Before using for the first time, add 100% ethanol to the Wash Buffers as shown on the side of the bottles.
Storage: Store the Lysis Buffer at 2 ~ 8 °C, protect from light. Store other components at room temperature (When using these buffers, be careful to avoid of contamination).
Note: Typically, 5~30 mg tissue is enough and best for most tissues. For tissues rich in RNA (such as liver, spleen and thymus), the amount of tissue should be reduced to no more than 5 mg/sample, to avoid of redundant 5S rRNA contamination.
Add equal volume of 100% ethanol to each volume of tissue homogenate, mix efficiently, and then, add to the spin column and centrifuge.
Wash with Wash Buffer 1 for twice, and then with Wash Buffer 2 for once.
Add Elution Buffer to the center of the column and incubate for minutes, then centrifuge to get the RNA.
Representative experimental results
Figure 1. RNA isolated from 100 mg mouse muscle (two parallel samples) using EZBioscience® Tissue RNA Purification Kit and TRIzol. M: 250bp DNA Ladder; Lanes 1 and 2: EZBioscience® Tissue RNA Purification Kit; Lanes 3 and 4: TRIzol; These results showed that more RNA can be isolated from the same amount of tissue by EZBioscience® Tissue RNA Purification Kit than TRIzol.
For detailed information, please check the product manual.