What is TRIzol-Based RNA Extraction?

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TRIzol-Based RNA Extraction is a technique applied for studies of gene expression, analysis of the etiology and disease progression, effects of given treatment, as well as designing the methods of diagnostic. TRIzol is proven to be an applicable material for RNA extraction from various biological samples in many studies. TRIzol is a combination of phenol, guanidine isothiocyanate and red dye. The all-in-one reagent offers excellent capability to lyse a variety of complex samples. TRIzol is easily scalable for accommodating very large inputs, recovers ample RNA yield and is also an effective inactivator of RNases, preventing degradation of samples during purification 1. Overall, TRIzol preserves the RNA quality, integrity and quantity, allowing it to double as a storage medium as well as an extraction solution.

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How does TRIzol-based RNA Extraction work with Covid-19 virus?

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TRIzol Reagent allows to perform sequential precipitation of RNA, DNA, and proteins from a single sample (Chomczynski, 1993). 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 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.

 

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Disadvantage of TRIzol-Based RNA Extraction.

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Although TRIzol is known to be a powerful reagent in itself, adding TRIzol to a sample is not a complete cure-all. One must consider the type of sample such as biological liquids, cells or solid tissue and the volume of TRIzol used. If the upstream processing is not well thought out, samples can result in incomplete lysis and DNA or phenol contamination.

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