TRIzol Reagent - User Guide

Oct 24, 2026

TRIzol™ Reagent user guide describes a ready-to-use chemistry for isolating high-quality total RNA, with parallel recovery of DNA and proteins, from human, animal, plant, yeast, or bacterial cell and tissue samples. It explains the monophasic phenol/guanidine-based workflow using phase separation with chloroform, then sequential precipitation with isopropanol and ethanol, followed by washing and resuspension. It also lists downstream applications for RNA, DNA, and protein, required materials, storage conditions, input sample limits, and essential safety handling instructions.

LanguageEnglish

CategoryHealthcare

FormatPDF

Pages4

Reading Time10 min

AccessFree to download

Product description

Contents and storage

Required materials not supplied

Table 1 Materials required for all isolations

Table 2 Materials required for RNA isolation

Table 3 Materials required for protein isolation

Input sample requirements

Procedural guidelines

USER GUIDE TRIzol ™ Reagent Catalog Numbers 15596026 and 15596018 Doc. Part No. 15596026. PPS Pub. No. MAN0001271 Rev. D WARNING! Read the Safety Data Sheets (SDSs) and follow the handling instructions. Wear appropriate protective eyewear, clothing, and gloves. Safety Data Sheets (SDSs) [are available from](are available from thermofisher.com/)[ thermofisher.com/](are available from thermofisher.com/)[ ](are available from thermofisher.com/)[support](support.)[.](support.) Product description Invitrogen™ TRIzol™ Reagent is a ready-to-use reagent, designed to isolate high quality total RNA (as well as DNA and proteins) from cell and tissue samples of human, animal, plant, yeast, or bacterial origin, within one hour. TRIzol™ Reagent is a monophasic solution of phenol, guanidine isothiocyanate, and other proprietary components which facilitate the isolation of a variety of RNA species of large or small molecular size. TRIzol™ Reagent maintains the integrity of the RNA due to highly effective inhibition of RNase activity while disrupting cells and dissolving cell components during sample homogenization. TRIzol™ Reagent allows for simultaneous processing of a large number of samples, and is an improvement to the single-step RNA isolation method. TRIzol™ Reagent allows users to perform sequential precipitation of RNA, DNA, and proteins from a single sample. After homogenizing the sample with TRIzol™ Reagent, chloroform is added, and the homogenate is allowed to separate into a clear upper aqueous layer (containing RNA), an interphase, and a red lower organic layer (containing the DNA and proteins) . RNA is precipitated from the aqueous layer with isopropanol. DNA is precipitated from the interphase/organic layer with ethanol. Protein is precipitated from the phenol-ethanol supernatant by isopropanol precipitation. The precipitated RNA, DNA, or protein is washed to remove impurities, and then resuspended for use in downstream applications. • Isolated RNA can be used in RT-PCR, Northern Blot analysis, Dot Blot hybridization, poly(A)+ selection, in vitro translation, RNase protection assay, and molecular cloning. • Isolated DNA can be used in PCR, Restriction Enzyme digestion, and Southern Blots. • Isolated protein can be used for Western Blots, recovery of some enzymatic activity, and some immunoprecipitation. For DNA isolation, see the TRIzol™ Reagent (DNA isolation) User Guide (Pub. No. MAN0016385) . TRIzol™ Reagent can also be used with Phasemaker™ Tubes (Cat. No. A33248) to isolate RNA. Phasemaker™ Tubes creates a solid barrier between the organic and aqueous phases of the TRIzol™ Reagent following sample homogenization which makes separation of phases easier. See the TRIzol™ Reagent and Phasemaker™ Tubes Complete System User Guide (Pub. No. MAN0016163) for the full protocol. TRIzol™ Reagent can also be used with the PureLink™ RNA Mini Kit (Cat. No. 12183018A ) which uses spin columns instead of ethanol precipitation to purify the RNA. For additional information, see the PureLink™ RNA Mini Kit User Guide (Pub. No. MAN0000406) . Contents and storage | Contents | Cat. No. 15596026 (100 reactions) | Cat. No. 15596018 (200 reactions) | Storage | | --- | --- | --- | --- | | TRIzol™
Reagent | 100 mL | 200 mL | 2–25°C | Required materials not supplied Unless otherwise indicated, all materials are available through [thermofisher.com](thermofisher.com). "MLS" indicates that the material is available from [fisherscientific.com](fisherscientific.com) or another major laboratory supplier. Table 1 Materials required for all isolations | Item | Source | | --- | --- | | Equipment | | | Centrifuge and rotor capable of reaching 12,000 × g and 4°C | MLS | | Tubes | | | Polypropylene microcentrifuge tubes | MLS | | Reagents | | | Chloroform | MLS | Table 2 Materials required for RNA isolation | Item | Source | | --- | --- | | Equipment | | | Water bath or heat block at 55–60°C | MLS | | Reagents | | |

FAQs

What does TRIzol™ Reagent isolate, and from which sample types?

TRIzol™ Reagent isolates high-quality total RNA, as well as DNA and proteins, from cell and tissue samples originating from human, animal, plant, yeast, or bacterial sources.

How are RNA, DNA, and proteins separated and precipitated in the workflow?

After homogenization, chloroform is added and the mixture separates into an upper aqueous phase (RNA), an interphase, and a red lower organic phase (DNA and proteins). RNA is precipitated from the aqueous layer with isopropanol, DNA from the interphase/organic layer with ethanol, and proteins from the phenol-ethanol supernatant by isopropanol precipitation.

What safety and handling precautions does the guide emphasize?

The guide instructs readers to read and follow Safety Data Sheets (SDSs), wear protective eyewear, clothing, and gloves, and follow procedural steps including aseptic technique and RNase contamination prevention. It also states to use cold reagent for RNase-rich samples and to change gloves frequently.

Oct 24, 2026

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