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| Product Name | TRI Reagent (Total RNA Isolation Solution) |
| Catalog No. | NATR-HMM-0111 |
| Description | A monophasic solution of phenol, guanidine isothiocyanate, and other proprietary components for the simultaneous isolation of total RNA, DNA, and protein from a single biological sample. This single-step method provides high yields of undegraded RNA suitable for demanding downstream applications. |
| Intended Use | Isolation of total RNA from tissues, cells, bacteria, yeast, plant material, and biological fluids for RT-PCR, Northern blotting, nuclease protection assays, microarray analysis, and RNA-seq library preparation. The reagent can also recover DNA and protein from the same sample. |
| Principle / Technology | The reagent maintains RNA integrity while disrupting cells and dissolving cellular components through a combination of phenol extraction and guanidine isothiocyanate denaturation. Addition of chloroform creates phase separation: RNA partitions into the aqueous phase, DNA remains at the interphase, and proteins partition into the organic phase. RNA is recovered by isopropanol precipitation from the aqueous phase. |
| Detection Method | UV spectrophotometry (A260/A280 for purity, A260/A230 for salt contamination); agarose or polyacrylamide gel electrophoresis with ethidium bromide or SYBR staining for RNA integrity; Bioanalyzer RIN analysis for quality assessment |
| Sample Type | Fresh or frozen cultured cells (adherent and suspension), fresh or frozen animal tissues, bacterial and yeast pellets, plant tissues, whole blood, plasma, serum, biological fluids |
| Performance Range / Specifications | Typical RNA yield from 10^7 cultured cells: 30–150 μg; from 100 mg tissue: 50–500 μg (tissue-dependent); A260/A280: 1.8–2.1; A260/A230: >2.0 |
| Sensitivity / LOD | Effective RNA isolation from as few as 200 cells; recovers low-abundance transcripts (<5 copies per cell) detectable by RT-qPCR |
| Specificity | Recovers total RNA including mRNA, rRNA, tRNA, miRNA, and other small RNAs; no size selection; maintains poly(A)+ RNA for oligo-dT-based purification; RNA integrity preserved for at least 24 hours at room temperature in TRI Reagent |
| Reaction Conditions / Protocol | Homogenize sample in 1 mL TRI Reagent per 10^7 cells or 100 mg tissue, incubate 5 minutes at room temperature, add 0.2 mL chloroform, shake vigorously for 15 seconds, incubate 3 minutes, centrifuge 12,000g for 15 minutes at 4°C, transfer aqueous phase to fresh tube, precipitate RNA with 0.5 mL isopropanol, incubate 10 minutes, centrifuge 12,000g for 10 minutes at 4°C, wash pellet with 75% ethanol, air dry, resuspend in nuclease-free water; optional DNA and protein recovery from organic phase and interphase |
| Components / Formulation | TRI Reagent (phenol, guanidine isothiocyanate, ammonium thiocyanate, sodium acetate, glycerol), nuclease-free water for RNA resuspension; chloroform, isopropanol, and 75% ethanol required but not supplied |
| Storage Conditions | 2–8°C protected from light in a tightly sealed, acid-resistant container; stable at room temperature for up to 2 weeks during processing |
| Shelf Life | 24 months from date of manufacture at recommended storage; after opening, use within 6 months |
| Package Specifications | 100 mL and 200 mL bottles (100 mL processes approximately 100 samples at 1 mL per sample) |
| Product Form | Monophasic liquid solution; red-brown colored due to phenol content |
| Quality Control | Each lot tested with HeLa cell RNA: yield ≥25 μg per 10^6 cells; A260/A280: 1.80–2.05; RIN score ≥8.5 as measured by Bioanalyzer; DNA and protein recovery verified from organic phase; functional testing by RT-PCR for GAPDH and β-actin |
| Key Features | Single-step total RNA, DNA, and protein isolation from one sample; no ultracentrifugation or cesium chloride gradients required; cost-effective with published protocol; scalable for samples from milligrams to grams; compatible with difficult tissues including fibrous and lipid-rich samples |
For research use only, not for clinical use.
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