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TRI Reagent® (Total RNA Isolation)
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TRI Reagent® LS (liquid samples) TRI Reagent®LS is a complete and ready to use reagent for the isolation of total RNA or the simultaneous isolation of RNA, DNA and proteins from liquid samples of human, animal, plant, yeast, bacterial and viral origin. TRI Reagent LS is the improved version of the popular single-step method of total RNA isolation (1, 2). This highly reliable technique performs well with small and large sample volumes and allows simultaneous processing of a large number of samples. TRI Reagent LS and the single-step method are subjects of the US patents 4,843,155 and 5,346,994, other patents pending. TRI Reagent LS combines phenol and guanidine thiocyanate in a mono-phase solution to facilitate the immediate and most effective inhibition of RNase activity. A biological sample is homogenized or lysed in TRI Reagent LS and the lysate is separated into aqueous and organic phases by bromochloropropane (cat. no. BP 151) or chloroform addition and centrifugation. RNA remains exclusively in the aqueous phase, DNA in the interphase and proteins in the organic phase. RNA is precipitated from the aqueous phase by the addition of isopropanol and the RNA pellet is washed with ethanol and solubilized. DNA and proteins are sequentially precipitated from the interphase and organic phase with ethanol and isopropanol, washed with ethanol and solubilized.STABILITY: TRI Reagent LS is stable at 25 C for at least two years from the date of purchase.   I. ISOLATION OF RNA TRI Reagent LS is the most effective method of RNA isolation. The entire procedure can be completed in 1 h and the recovery of undegraded mRNAs is 30-150% greater than with other methods of RNA isolation. TRI Reagent LS isolates a whole spectrum of RNA molecules rarely observed in RNA isolated by other methods. Typically, the column-based methods artificially change the mRNA composition. TRI Reagent LS isolates RNA from diverse biological material including animal and plant tissues rich in polysaccharides and proteoglycans. The isolated RNA and can be used for northern analysis, dot blot hybridization, poly A+ selection, in vitro translation, RNase protection assay, molecular cloning and RT-PCR. Simultaneous extraction of nearly 100% of the genomic DNA allows for normalization of the results of gene expression studies to genomic DNA instead of the more variable total RNA or tissue weight. * Patents granted to Hoffmann LaRoche Corporation. PROTOCOL Reagents required, but not supplied: chloroform or 1-bromo-3-chloropropane (BCP, cat. no. BP 151), isopropanol and ethanol. We recommend the use of disposable polypropylene tubes provided by Molecular Research Center, Inc. Tubes from other suppliers should be tested to ensure integrity during centrifugation at 12,000 g with TRI Reagent LS. The protocol includes the following steps: 1.  HOMOGENIZATION:   0.75 ml TRI Reagent LS + 0.25 ml sample. 2.  PHASE SEPARATION:   homogenate + 0.1 ml BCP or 0.2 ml chloroform. 3.  RNA PRECIPITATION:   aqueous phase + 0.5 ml isopropanol. 4.  RNA WASH:   1 ml 75% ethanol. 5.  RNA SOLUBILIZATION:   FORMAzol®, 0.5% SDS, or water.Ordering Information: TRI Reagent®LS (100 ml)                                                                                     Cat. TS120100  TRI Reagent®LS (200 ml)                                                                                     Cat. TS120200 Store at RT                                                                                                 For research use only     Details...

Total RNA Mini Kit (Blood) The total RNA Mini Kit (Blood and cultured cell) is specially designed for purification of total RNA from cultured cells and fresh whole human blood. This method uses detergents and a chaotropic salt to lyse cells and inactivate RNase then RNA in chaotropic salt is bonded to the glass fiber matrix of the column. After washing off the contaminants, the purified RNA is eluted by RNase-free water. Sample: blood, cultured cells, body fluids and other animal tissues Sample size: 5×106 animal cultured cells, 1×109 bacterial cells, or 300μl blood Format: spin columns Operation: centrifuge/vacuum Binding capacity: up to 60μg Operation time: 30minutes Applications: - RT-PCR - Real-time RT-PCR - Northern blotting - Primer extension - RNase protection assays - mRNA selection - cDNA synthesis  Ordering Information: Total RNA Mini Kit (Blood) 50 rxns                                                                          Cat. RB050 Total RNA Mini Kit (Blood) 100 rxns                                                                          Cat. RB100 Store at RT                                                                                                 For research use only   Details...

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