Life Technologies
TRIzol
15596-026
We use TRIzol to extract RNA from various cells and tissues for PCR amplification. The general aim is to study changes in gene expression. I started using TRIzol in graduate school and still use it because I find that it's less expensive per use than prep kits. It works well for our applications.
RNA isolation
Mammalian cells, tissues
I don't deviate from the easy-to-follow protocol included with the reagent. In short, the starting material is lysed in TRIzol and the RNA is extracted to an upper aqueous phase by addition of cholorform. The RNA is precipitated/washed with alcohol and solubilized in water. TRIzol is nearly fool-proof because it's capable of robust lysis and RNAse inactivation. It's versatile in that protein can be collected from the organic phase and DNA from the interface.
Go for purity rather than quantity when collecting the desired phase
My undergraduate researchers isolated RNA from tissue (A260/280 ~ 1.9), amplified targets using PCR, and ran the amplicons on a gel for imaging.
None
Excellent results
Hazardous chemicals
A time-tested, versatile, and economical method for RNA isolation. The only drawbacks are that it's more time-consuming than spin column-based kits and involves hazardous wastes.