Purification of RNA Using TRIzol (TRI Reagent)
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- 1 June 2010
- journal article
- Published by Cold Spring Harbor Laboratory in Cold Spring Harbor Protocols
- Vol. 2010 (6)
- https://doi.org/10.1101/pdb.prot5439
Abstract
INTRODUCTION: TRIzol solubilization and extraction is a relatively recently developed general method for deproteinizing RNA. This method is particularly advantageous in situations where cells or tissues are enriched for endogenous RNases or when separation of cytoplasmic RNA from nuclear RNA is impractical. TRIzol (or TRI Reagent) is a monophasic solution of phenol and guanidinium isothiocyanate that simultaneously solubilizes biological material and denatures protein. After solubilization, the addition of chloroform causes phase separation (much like extraction with phenol:chloroform:isoamyl alcohol), where protein is extracted to the organic phase, DNA resolves at the interface, and RNA remains in the aqueous phase. Therefore, RNA, DNA, and protein can be purified from a single sample (hence, the name TRIzol). TRIzol extraction is also an effective method for isolating small RNAs, such as microRNAs, piwi-associated RNAs, or endogeneous, small interfering RNAs. However, TRIzol is expensive and RNA pellets can be difficult to resuspend. Thus, the use of TRIzol is not recommend when regular phenol extraction is practical.Keywords
This publication has 4 references indexed in Scilit:
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- Preparation of Cytoplasmic and Nuclear RNA from Tissue Culture CellsCold Spring Harbor Protocols, 2010
- Ethanol Precipitation of RNA and the Use of CarriersCold Spring Harbor Protocols, 2010
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