A reagent for the single-step simultaneous isolation of RNA, DNA and proteins from cell and tissue samples.
This report describes a new method for simultaneous isolation of RNA, DNA and proteins from cell and tissue samples. The method is based on the use of a reagent containing phenol and guanidine thiocyanate. A biological sample is homogenized in the reagent and the simultaneous isolation of RNA, DNA and proteins is accomplished in a single step by a liquid-phase separation. The isolation of RNA can be completed in about 1 h, and DNA and proteins in about 3 h. The simultaneously isolated RNA, DNA and proteins are ready for Northern, Southern and Western blotting. The complete recovery of DNA from samples used for the RNA and protein isolation makes it possible to normalize the results of gene expression studies based on DNA content instead of on the more variable total RNA, protein content or tissue weight.
Chomczynski
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College of Medicine, University of Cincinnati.
D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D015153:Blotting, Western; D001940:Breast / Q000737:chemistry; D004247:DNA / Q000302:isolation & purification*; D006146:Guanidines; D006801:Humans; D007202:Indicators and Reagents; D007768:Lactalbumin / Q000235:genetics; D008321:Mammary Glands, Animal / Q000737:chemistry; D019800:Phenol; D010636:Phenols; D011506:Proteins / Q000302:isolation & purification*; D012313:RNA / Q000302:isolation & purification*; D012333:RNA, Messenger / Q000302:isolation & purification; D051381:Rats; D013861:Thiocyanates; D013950:Thymus Gland / Q000737:chemistry; D013954:Thyroglobulin / Q000235:genetics; D013997:Time Factors
D006146:Guanidines; D007202:Indicators and Reagents; D010636:Phenols; D011506:Proteins; D012333:RNA, Messenger; D013861:Thiocyanates; D019800:Phenol; C054436:guanidine thiocyanate; D012313:RNA; D004247:DNA; D013954:Thyroglobulin; D007768:Lactalbumin