Extraction+using+TRIzol

__**TRIzol Extraction **__


 * Technique **

Extraction is a scientific technique that is described as a process of separation of a substance from a mixture or matrix. TRIzol is a reagent used in the extraction of DNA, RNA and proteins. The extraction technique, when using TRIzol, can be explained by brief steps as follows. First, the sample has TRIzol added and is left to homogenize. Chloroform is added and after sample is mixed well by inversion, phase separation occurs by centrifuging. With the RNA of the sample being in the top phase, it is removed and isopropyl alcohol is added. Incubation and another round of centrifuging follows. The RNA pellet is washed with 75% ethanol and centrifuged one last time resulting in the extracted RNA pellet [1].


 * Purpose **

The ”TRI” in the reagents name stands for the total RNA isolation as the DNA and proteins are isolated sequentially [2]. With that being known, the purpose of extraction using TRIzol is to isolate total RNA from a sample. RNA is not the only substance isolated. By only using one sample, RNA, DNA and proteins can be isolated and purified. The reagent is a mixture of phenol and guanidine isothiocyanate. A red dye is also a component of the reagent, which has the purpose of detection of organic phase. The detection is considered easy because the red dye is non interactive with nucleic acids.


 * Origins **

Piotr Chomczynski was the scientist who invented TRIzol.


 * Research **

The first research article, “Gene silencing of IL-12 in dendritic cells inhibits autoimmune arthritis”[3], utilized TRIzol to extract the total RNA from dendritic cells of mice. This technique was important in this research because the RNA was used to synthesize the cDNA. This then was followed by PCR to show the results of gene expression of IL-12 when IL-12 shRNA is silenced.


 * References **

[1] TRIzol® Reagent and TRIzol® LS Reagent [] [2] Kirkland, P., Busby, J., Stevens, S., & Maupin-Furlow, J. (2006). Trizol-based method for sample preparation and isoelectric focusing of halophilic proteins. Analytical Biochemistry, 351, 254-259. [3] Li, R., Zheng, X., Popov, I., Zhang, X., Wang, H., Suzuki, M., Necochea-Campion, R., & French, P. (2012). Gene silencing of il-12 in dendritic cells inhibits autoimmune arthritis. Journal of Translational Medicine, 10(19).