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Purification of RNA using TRIzol (TRI reagent)
Cold Spring Harbor Protocols
|June 3, 2010
Summary
TRIzol extraction is a method for deproteinizing RNA, useful for samples with high RNase activity or when separating RNA types is difficult. It isolates RNA, DNA, and protein but is costly and challenging for RNA resuspension.
Area of Science:
- Biochemistry
- Molecular Biology
- Genomics
Background:
- TRIzol reagent is a phenol and guanidinium isothiocyanate solution.
- It is used for deproteinizing RNA and solubilizing biological samples.
- This method is advantageous for samples with high endogenous RNase activity or when separating cytoplasmic from nuclear RNA is impractical.
Purpose of the Study:
- To describe the TRIzol method for RNA, DNA, and protein extraction.
- To highlight its utility in isolating small RNAs.
- To outline the limitations of the TRIzol method.
Main Methods:
- TRIzol reagent solubilizes biological material and denatures proteins.
- Chloroform addition induces phase separation, isolating RNA in the aqueous phase.
- This method allows simultaneous purification of RNA, DNA, and protein from a single sample.
Main Results:
- TRIzol extraction effectively isolates RNA, DNA, and protein.
- It is also effective for isolating small RNAs, including microRNAs and piwi-associated RNAs.
- Challenges include the high cost of TRIzol and difficulties in resuspending RNA pellets.
Conclusions:
- TRIzol is a versatile method for nucleic acid and protein extraction, particularly useful for specific biological samples.
- Its effectiveness in isolating small RNAs makes it valuable in molecular biology research.
- However, its cost and RNA resuspension issues limit its application when simpler methods suffice.
