• Rapid, single-step RNA extraction from cells and tissues
• Preserves RNA integrity and minimizes degradation
• Suitable for samples from various species
• Compatible with multiple downstream applications
• Allows simultaneous processing of multiple samples
• Simple protocol, completed within one hour

TRIpure Reagent is an improved single-phase solution of phenol and guanidine isothiocyanate for rapid, efficient extraction of total RNA from cells and tissues. It ensures RNA integrity by minimizing degradation during sample lysis and homogenization. After adding chloroform and centrifuging, the sample separates into aqueous and organic phases. RNA is located in the aqueous phase. After collecting the upper aqueous phase, RNA can be precipitated with isopropanol. After removing the aqueous phase, DNA and proteins in the sample can also be precipitated sequentially. Ethanol precipitation can isolate DNA from the intermediate phase and adding isopropanol to the organic phase precipitates proteins. Co-purification of DNA is useful for standardizing RNA yield across samples.

This method works well for isolating RNA from small amounts of tissue (50-100 mg) and cells (5×10⁶) as well as large amounts of tissue (≥1 g) and cells (>10⁷), regardless of whether the tissue is from human, animal, plant, or bacterial origin. The simplicity of the TRIpure reagent allows for processing multiple samples simultaneously. All procedures can be completed within one hour. The total RNA extracted with TRIpure is free of DNA and protein contamination, making it suitable for RNA blotting, dot blot hybridization, poly(A)+ selection, in vitro translation, RNase protection assay, and molecular cloning. For PCR, if both primers are located within a single exon, it is recommended to treat the extracted total RNA with DNase I before amplification.

The TRIpure reagent facilitates the extraction of various RNA molecules of different molecular weights from different species. For example, RNA extracted from rat liver, when analyzed by agarose gel electrophoresis and stained with ethidium bromide, shows numerous discontinuous high-molecular-weight bands between 7 kb and 15 kb (mRNA and hnRNA components), two dominant ribosomal RNA bands at ~5 kb (28S) and ~2 kb (18S), and low-molecular-weight RNA between 0.1 and 0.3 kb (tRNA, 5S). The A260/A280 ratio of the extracted RNA diluted in TE is ≥ 1.8.

• Chloroform
• Isopropanol
• 75% Ethanol
• RNase-free water or 0.5% SDS solution

1. Homogenize your tissue or cells in TRIpure reagent.
2. Transfer the homogenized sample to a clean tube.
3. Add chloroform, shake, and centrifuge to separate the mixture into phases.
4. Collect the upper aqueous phase, which contains RNA.
5. Precipitate RNA by adding isopropanol and centrifuge again.
6. Remove the supernatant and wash the RNA pellet with 75% ethanol.
7. Air-dry or vacuum-dry the RNA pellet briefly.
8. Resuspend the RNA pellet in RNase-free water or 0.5% SDS solution.
9. Store the purified RNA as needed for downstream applications.