Isolation of RNA bacterial cell

TRIzol  reagent is an acid-guanidinium-phenol based reagent ideally designed for the extraction of RNA (as well as DNA and protein) from various biological sample inputs. The low pH (acidic) of TRIzol controls to separate RNA from DNA and protein, while a high pH can cause RNA and DNA to be isolated together. The guanidinium salt serves as a chaotropic agent to denature proteins and the phenol (commonly indicated as a pink color) is an organic compound also used to extract nucleic acids and proteins)

After solubilization and homogenization of samples in TRIzol, the RNA, DNA and protein are differentially extracted by the addition of a phase separation reagent (chloroform, BCP or BAN). The solution separate-the RNA away from DNA and protein into different layers. An upper, clear aqueous phase mainly contains RNA, and the middle interphase and lower organic phase contains DNA, proteins and lipids 3. Subsequently, the RNA in the upper aqueous phase is then collected by alcohol-based precipitation Total RNA is then recovered by precipitation with isopropanol. RNase enzymes can be inactivated by including diethyl pyrrocarbonate (DEPC).

Total RNA is isolated and separated from DNA and protein after extraction with a solution called as Trizol. Trizol is an acidic solution containing guanidinium thiocyanate (GITC), phenol and chloroform. GITC irreversibly denatures proteins and RNases. This is followed by centrifugation.  TRIzol works by maintaining RNA integrity during tissue homogenisation, while at the same time disrupting and breaking down cells and cell components.