Isolation of RNA Using Trizol

  • Trizol is a very popular reagent among researchers for RNA isolation due to the simplicity and reliability of the isolation procedure with high yields of good quality RNA suitable for most downstream applications. 
  • Trizol is a Ready-to-use, monophasic solution of guanidinium isothiocyanate and phenol and some other proprietary components. 
  • It can be used for the isolation of total RNA from cells and tissues from a variety of organisms including bacteria, yeast, fungi, animals, plants etc. In addition to RNA, Trizol-based methods also allow purification of protein and DNA from the same sample. 
  • Several samples can be processed simultaneously but require skills and practice to perform isolation procedures. In addition, Trizol use requires appropriate safety measures as its constituents such as guanidinium isothiocyanate and phenol, are extremely harmful.
  • Although Trizol itself is very efficient at solubilizing and isolating RNA from cell culture, additional reagents and steps can be required to process tissues, and difficult to solubilize cells (yeast cells, bacterial cells due to cell wall). 
  • Trizol quickly denatures proteins including RNases upon lysis. It is extremely important to inactivate RNases as soon as cells are lysed and all cellular components are in solution and free to interact. 
  • Any delay in the inactivation of RNases results in the degradation of RNA. Therefore, tissues that are enriched with RNases must be processed carefully to avoid RNA degradation during solubilization. Special reagents like RNA later can be used to protect RNA from RNase-mediated degradation in RNase-rich tissues.
  • Trizol purified RNA may contain a very low amount of genomic DNA contamination. If the RNA is to be used for applications like quantitative PCR which can not tolerate any genomic DNA contamination (primers designed within the same exons), a DNase I treatment is recommended to remove traces of DNA from the RNA samples.

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