Using Tri Reagent to Extract Plant DNA

D. KIM dkim at NMSU.Edu
Wed Jun 3 18:35:46 EST 1998


The Tri-Reagent is a variant of the "Acid Guanidinium-Phenol-Chloroform"
RNA extraction method, in which guanidine thiocyanate is used to disrupt
cells and denature proteins, while phenol at pH <5 extracts protein and
DNA into the organic phase/interphase.  RNA is then recovered from the
aqueous phase.

If the phenol is then re-equilibrated to neutrality, the DNA is supposed
to partition into the aqueous phase for another extraction.

I forget how the protein is supposed to be recovered from phenol (I always
throw away the protein :))

The original AGPC method was reported in ANalytical Biochemistry in
(maybe) 1985 by P. Chomczynski and N. Sacchi (I may have the spelling
wrong), and was quickly converted into a kit method, to the detriment of
research budgets everywhere.

Daniel Kim

Mary_Wood at BAYLOR.EDU wrote:
: I've been TRYING to use a product called Tri-Reagent to extract genomic DNA from 
: tobacco plants.  Supposedly the DNA is in the interphase and phenol phase of the 
: mixture after removal of RNA in the aqueous phase.  But I am getting very little 
: DNA -- about 200 ng from 100 mg of plant tissue after homogenizing in liquid 
: nitrogen.  Has anyone used this product to extract genomic plant DNA?  How did 
: you do it? 




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