Isolation of deoxyribonucleic acid and ribosomal ribonucleic acid from bacteria.
1. Nucleic acids were released from Escherichia coli by lysing with tri-iso-propylnaphthalene sulphonate and 4-aminosalicylate and then extracting with a phenol-cresol mixture. 2. Nucleic acids were similarly released from Bacillus subtilis after initial treatment with lysozyme. 3. DNA was sedimented after careful precipitation with m-cresol or 2-butoxyethanol (0.1-0.12vol.) in the presence of 20% sodium benzoate. 4. Contaminating ribosomal RNA was removed by precipitation in the presence of 4m-sodium chloride or by extracting DNA with an acetate-butyrate mixture, in which RNA is insoluble. 5. The DNA from B. subtilis has a transforming ability of 0.3-0.6% for the tryptophan marker. 6. Ribosomal RNA was then precipitated with rapidly labelled RNA by the addition of an equal volume of 2-butoxyethanol. 7. There was good separation of the nucleic acids from protein and polysaccharides.
D001412:Bacillus subtilis / Q000032:analysis* / Q000166:cytology; D011232:Chemical Precipitation; D004269:DNA, Bacterial / Q000032:analysis*; D004926:Escherichia coli / Q000032:analysis* / Q000166:cytology; D006358:Hot Temperature; D009691:Nucleic Acid Denaturation; D012329:RNA, Bacterial / Q000032:analysis*; D012270:Ribosomes / Q000032:analysis*; D014461:Ultracentrifugation
D004269:DNA, Bacterial; D012329:RNA, Bacterial