Description
Product Introduction:TRzol Reagent is a reagent designed for the direct extraction of total RNA from cells or tissues. It maintains the integrity of RNA during the disruption and lysis of cells. Upon the addition of a chloroform substitute/chloroform and subsequent centrifugation, the sample separates into an aqueous phase, an interphase, and an organic phase. RNA is present in the aqueous phase. After collecting the upper aqueous phase, RNA can be recovered by isopropanol precipitation. This method is effective for both small and large amounts of tissues and cells, regardless of whether they are from humans, animals, plants, or bacteria. The simplicity of TRzol Reagent allows for the simultaneous processing of multiple samples. All operations can be completed within one hour. Total RNA extracted with TRzol is free from DNA and protein contamination and can be used for RNA blotting analysis, dot hybridization, poly(A)+ selection, in vitro translation, RNase protection analysis, and molecular cloning. When used for PCR, it is recommended to treat the extracted total RNA with amplification-grade DNase I if both primers are located within a single exon. TRzol Reagent facilitates the precipitation of various types of RNA with different molecular weights across species. For example, RNA extracted from rat liver, when subjected to agarose gel electrophoresis and stained with ethidium bromide, shows numerous discontinuous high molecular weight bands between 7kb and 15kb (mRNA and hnRNA components), two major ribosomal bands with molecular weights of approximately 5kb (28S) and 2kb (18S), and low molecular weight RNA between 0.1 and 0.3kb (tRNA, 5S). When the extracted RNA is diluted with TE, its A260/A280 ratio is ≥1.8.Product component:T669996Component100 mlStorageT669996ATRzol100 ml2-8℃. Store in the dark.T669996B Chloroform substitute 10 ml2-8℃. Store in the dark. Precautions:1. Isolation of RNA samples from small amounts of tissue (1-10mg) or cells (10²-10⁴ cells): Add 800µl of TRzol to the tissue or cells. After the sample is lysed, add the chloroform substitute and proceed with the extraction steps. Before precipitating RNA with isopropanol, add 5-10µg of RNase-free glycogen as a carrier for the aqueous phase. To reduce viscosity, pass the sample through a 26-gauge needle twice before adding chloroform to shear genomic DNA. Glycogen will remain in the aqueous phase and co-precipitate with RNA. It will not inhibit the synthesis of the first strand in reverse transcription reactions or PCR until it is concentrated to 4mg/ml.2. Sample storage: The sample can be stored at -60℃ or -70℃ for at least one month before homogenization and addition of the chloroform substitute (R749970). RNA precipitate dissolved in 75% ethanol can be stored at 2-8℃ for at least one week and at -5 to -20℃ for at least one year.3. Safety: When extracting RNA with TRzol, wear gloves and eye protection. Avoid contact with skin and clothing. Perform the operation in a chemical fume hood. Avoid inhalation through the respiratory tract. Unless otherwise specified, maintain the room temperature between 15-30℃