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Guanidinium thiocyanate (GTC) or guanidinium isothiocyanate (GITC) is a chemical compound used as a general protein denaturant, being a chaotropic agent, although it is most commonly used as a nucleic acid protector in the extraction of DNA and RNA from cells. [1] GITC may also be recognized as guanidine thiocyanate.
TRIzol reagent contains guanidinium thiocyanate and phenol.. TRIzol is a widely used [1] chemical solution used in the extraction of DNA, RNA, and proteins from cells. The solution was initially used and published by Piotr Chomczyński and Nicoletta Sacchi in 1987.
RNA partitions in the aqueous phase, while proteins and DNA partition into the organic/interphase (left). The RNA is then precipitated in an alcohol (right). Acid guanidinium thiocyanate-phenol-chloroform extraction (abbreviated AGPC) is a liquid–liquid extraction technique in biochemistry and molecular biology.
According to the original Boom method, the chaotropic guanidinium salt employed is preferably guanidinium thiocyanate (GuSCN). According to the chaotropic effect, in the presence of the chaotropic agent, hydration water of nucleic acids are taken from the phosphodiester bond of the phosphate group of the backbone of a nucleic acid.
This was later improved using guanidinium thiocyanate or guanidinium hydrochloride as the chaotropic agent. [4] For ease of handling, the use of glass beads was later changed to silica columns. And to enable use of automated extraction instruments, there was development of silica-coated paramagnetic beads, more commonly referred to as "magnetic ...
A chaotropic agent is a substance which disrupts the structure of, and denatures, macromolecules such as proteins and nucleic acids (e.g. DNA and RNA).Chaotropic solutes increase the entropy of the system by interfering with intermolecular interactions mediated by non-covalent forces such as hydrogen bonds, van der Waals forces, and hydrophobic effects.
In this example total RNA is isolated (both nuclear and cytoplasmic) by guanidinium thiocyanate-phenol-chloroform extraction (e.g. Trizol) which isolates most RNA (whereas column methods have a cut off of 200 nucleotides) and if done correctly has a better purity.
Several methods are used in molecular biology to isolate RNA from samples, the most common of these is guanidinium thiocyanate-phenol-chloroform extraction. [2] [3]. Usually, the phenol-chloroform solution used for RNA extraction has lower pH, this aids in separating DNA from RNA and leads to a more pure RNA preparation.