RNA extraction from blood TRIzol Method with Demo
RNA extraction is simply the process of extraction of purified RNA from the source. The source can be anything blood sample bacterial cell animal cell or plant cell. Extraction of RNA is a comparatively sensitive process because RNA is not as stable as DNA as it is single stranded. So methods of extraction of DNA cannot be applied directly for isolation of RNA because it is much more sensitive. Basic principle is the breakdown of cell wall (in case of bacteria and plant cells) and cell membrane, removal of all other contaminants (protein, lipids, DNA and carbohydrates for eukaryotic cells), concentration of RNA and analysis.
TRIzol™ Reagent allows to perform sequential precipitation of RNA, DNA, and proteins from a single sample (Chomczynski, 1993).
After homogenizing the sample with TRIzol™ Reagent, chloroform is added, and the homogenate is allowed to separate into a clear upper aqueous layer (containing RNA), an interphase, and a red lower organic layer (containing the DNA and proteins).
RNA is precipitated from the aqueous layer with isopropanol. DNA is precipitated from the interphase/organic layer with ethanol. Protein is precipitated from the phenol-ethanol supernatant by isopropanol precipitation.
The precipitated RNA, DNA, or protein is washed to remove impurities, and then resuspended for use in downstream applications.