The preparation of ribosomal ribonucleic acid from whole bacteria.
1. A simple method for the preparation of ribonuclease-free ribosomal RNA is described in which ribonuclease-deficient bacteria are treated with acetone and the RNA is extracted with phenol and purified by precipitating it with potassium acetate. The treatment with acetone appears to render the cell wall permeable to RNA but not to DNA during the extraction with phenol. The method thus avoids the need to disrupt the bacteria and greatly simplifies the subsequent purification. 2. The method has been used successfully with ribonuclease-deficient strains of Escherichia coli, Pseudomonas fluorescens and Staphylococcus epidermidis. The recovered purified RNA accounts for about 70% of the total ribosomal RNA and shows the normal sedimentation pattern of the 16s and 23s components in the analytical centrifuge.
D000085:Acetates; D000096:Acetone; D004269:DNA, Bacterial; D004926:Escherichia coli; D000431:Ethanol; D006358:Hot Temperature; D008722:Methods; D010636:Phenols; D011549:Pseudomonas; D012329:RNA, Bacterial / Q000032:analysis*; D012270:Ribosomes / Q000032:analysis*; D013210:Staphylococcus; D013997:Time Factors; D014461:Ultracentrifugation
D000085:Acetates; D004269:DNA, Bacterial; D010636:Phenols; D012329:RNA, Bacterial; D000096:Acetone; D000431:Ethanol