Uses
TRI Reagent
? has been used for isolating RNA from various types of cells or tissues. It may also be used for processing tissues, cells cultured in monolayer, or cell pellets.
Uses
TRI Reagent has been used:
- in the isolation of total RNA from various tissue samples and cells
- for the lysis of monocytes
- to isolate cytoplasmic RNA from SARS-CoV infected Vero cells
- to isolate total RNA from MDMs (monocyte-derived macrophages) infected with influenza A (H5N1) virus
- to prepare a viral lysate of SARS-CoV culture for the creation of a clone encoding the S glycoprotein in the development of SARS-CoV vaccine
The RNA, DNA and protein that are isolated using TRI reagent
? can further be used for downstream applications, such as cloning, PCR, RT-PCR, Northern blots, mRNA isolation,
in vitro translation, RNase protection assay, restriction enzyme digestion, Southern blots, SDS-PAGE and western blots.
General Description
TRI Reagent LS is a quick and convenient reagent for use in the simultaneous isolation of RNA, DNA, and protein from liquid samples of human, animal, plant, yeast, bacterial, and viral origin.
A convenient single-step liquid phase separation results in the simultaneous isolation of RNA, DNA, and protein. This procedure is an adaptation of the singlestep method reported by Chomczynski and Sacchi for total RNA isolation, and permits fast and efficient processing of liquid samples. TRI Reagent LS performs well with large or small sample volumes, and many samples can be simultaneously extracted.
TRI Reagent LS is a mixture of guanidine thiocyanate and phenol in a monophase solution. When a biological sample is homogenized or lysed with it, and chloroform or 1-bromo-3-chloropropane is added, the mixture separates into 3 phases: an aqueous phase containing RNA, the interphase containing DNA, and an organic phase containing proteins. Each component can then be isolated after separating the phases. 0.75 ml of TRI Reagent LS processes 0.25 ml of a liquid sample such as amniotic fluid.
This is one of the most effective methods for isolating total RNA from fresh samples in only one hour. The procedure is very effective for isolating RNA molecules of all types from 0.1-15 kb in length. The resulting RNA is intact with little or no contaminating DNA and protein.
