Comments
The simultaneous isolation of RNA, DNA and proteins from biological samples has been introduced by Chomczynski (1) and improved by others. Chomczynski's method is based on the isolation of non-degraded nucleic acids (RNA and DNA) with guanidinium salts and phenol (e. g. ref. 2-5).TRItidy G™ is a reagent, based on the Chomczynski method (1), with additional modifications to improve the purity of the RNA, DNA and proteins. First, the RNA will be selectively retained in the aqueous phase during the acidic GuaSCN/Phenol extraction, while DNA and proteins stay in the organic phase and interphase, respectively (6). The DNA is isolated from the interphase/organic phase by a simple ethanol precipitation and proteins can be isolated from the remaining organic phase.
Literature
(1) Chomczynski, P. (1993) BioTechniques 15, 532-537A single-step purification method for RNA, DNA and proteins from cell and tissue samples. (2) Cox, R.A. (1968) Methods Enzymol. 12 Part B, [103a] 120-129. The use of guanidinium chloride in the isolation of nucleic acids. (3) Kirby, K.S. (1968) Methods Enzymol. 12 Part B, [98] 87-99. Isolation of nucleic acids with phenolic solvents. (4) MacDonald, R.J. et al. (1987) Methods Enzymol. 152, 219-227. Isolation of RNA with guanidinium salts. (5) Feramisco, J.R. et al. (1982) J. Biol. Chem. 257, 11024-11031. Modification of the GTC-Method for the RNA isolation according to Chirgwin. (6) Wallace, D.M. (1987) Methods Enzymol. 152, 33-41. Large and small scale phenol extraction. (7) Chomczynski, P. & Mackey, K. (1995) Anal. Biochem. 225, 163-164. Substitution of chloroform by bromochloropropane in the Single-step method of RNA-isolation (8) Chomczynski, P. & Mackey, K. (1995) BioTechniques 19, 942-945. Modification of the TRI Reagent procedure for isolation of RNA from polysaccharide- and proteoglycan-rich sources. (9) Monstein, H.-J. et al. (1995) BioTechniques 19, 340-344. RNA extraction from gastrointestinal tract and pancreas by a modified Chomczynski and Sacchi method.
