Hi all,
I know Guanidium Thiocyanate can lyse the cell in the RNA extraction process, but how? Does it a reagent and interact with phospholipids so in this way lyse the cell or just by protein denaturing?
Guanidine thiocyanate is a potent and interfering with the hydrogen bond network in aqueous solutions, it has a destabilizing effect on proteins. RNAases are proteins which are the most stable proteins. You need strong denaturants such as Guanidium. This will destroy the RNA degrading enzyme so that you could get good quality RNA.
It is commonly used in cells and the lysis processes in virus particles to extract nucleic acids, as it denatures RNAse and DNAse enzymes that would otherwise damage the extract. In the recent COVID-19 pandemic, this has found extensive application among researchers in the extraction of nucleic acid from SARS-Cov-2 virus for vaccine research and development.
Guanidine thiocyanate act by proteins denaturation, its due e to the combination of the two components : guanidinium cation and thiocyanate anion, which are chaotropic agents.
I have a question related to guanidine thiocyanate. As you say it denature the proteins, I wonder if it is possible to perform western blot using such lysates or if the protein are degraded ?
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